bluuebunny/crossref_metadata_2025 huggingface dataset:
Screening for familial disease presence in first-degree relatives of Behçet’s disease patients: Is measurement of common femoral vein wall thickness valuable for the diagnosis?
Authors: Boncukcuoğlu Ayşe Elif , Topal Erhan , Ergelen Rabia , Kaş Dilara , Alkan Elifnur , Kaya Gamzenur , Girgin Şevket , Ergun Tülin , Direskeneli Haner , Alibaz-Oner Fatma
Published: August 2024

Objectives

We aimed to assess first-degree relatives (FDRs) of Behçet’s disease (BD) patients for the presence of clinical symptoms and signs of BD and evaluate common femoral vein (CFV) wall thickness measurement for the diagnosis.

Methods

Patients with BD (n = 129) and FDRs (n = 230) of these patients were included. FDRs were questioned in terms of BD symptoms by phone. Pathergy test and CFV wall thickness measurement were performed among 111 FDRs who accepted the clinical assessment. Clinical assessment groups were classified according to the criteria sets for BD. FDRs who did not meet the criteria sets and had at least one clinical finding in addition to oral aphthae (OA) were categorized as the ‘suspected BD’.

Results

: We observed increased frequency of isolated BD manifestations in FDRs. Ten FDRs were diagnosed with BD during clinical evaluation. A significantly increased CFV wall thickness was observed in FDRs of BD patients fulfilling diagnostic BD criteria (P < .001 for both sides) and also in those with suspected BD group (P < .05 for both sides). The presence of OA, genital ulcer, folliculitis, or erythema nodosum was associated with increased CFV wall thickness (P < .05).

Conclusion

Our results suggest that the CFV wall thickness measurement can be used in the diagnosis of familial BD.
Enhancing prognostication and treatment response evaluation in primary CNS lymphoma with 18F-FDG-PET/CT
Authors: Oh Minyoung , Cho Hyungwoo , Park Ji Eun , Kim Ho Sung , Go Heounjeong , Park Chan-Sik , Lee Sang-wook , Song Sang Woo , Kim Young-Hoon , Cho Young Hyun , Hong Seok Ho , Kim Jeong Hoon , Lee Dong Yun , Ryu Jin-Sook , Yoon Dok Hyun , Kim Jae Seung
Published: August 2024

Background

The role of 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG-PET/CT) in the prognostication and response evaluation of primary central nervous system lymphoma (PCNSL) remains inadequately defined.

Methods

We conducted a retrospective analysis of 268 consecutive newly diagnosed patients with PCNSL between 2006 and 2020. Of these patients, 105 and 110 patients were included to evaluate the prognostic value of baseline and post-treatment 18F-FDG-PET/CT scans, respectively. Tumor uptake was considered positive when it exceeded that of the contralateral brain upon visual assessment. Quantitative analysis of baseline 18F-FDG-PET/CT included measurement of the maximal standardized uptake value (SUVmax), total metabolic tumor volume (TMTV), and total lesion glycolysis (TLG).

Results

The median age of the 268 patients was 62 years (range: 17–85), with 55% being male. The median progression-free survival (PFS) was 24.5 months (95% CI: 19.9–29.1), and the median overall survival (OS) was 34.5 months (95% CI: 22.9–46.1). The average SUVmax was 15.3 ± 5.7 and the mean TMTV and TLG were 12.6 ± 13.9 cm3 and 135.0 ± 152.7 g, respectively. Patients with a baseline TMTV ≥ 17.0 cm3 had significantly shorter OS (12.5 vs 74.0 months, P = .011). Post-treatment metabolic response by 18F-FDG-PET/CT significantly predicted PFS (median: 10.5 vs 46.0 months, P = .001) and OS (median: 21.0 vs 62.0 months, P = .002), whereas anatomic response by contrast-enhanced MRI showed no statistically significant differences in PFS (P = .130) or OS (P = .540).

Conclusion

Baseline TMTV and post-treatment metabolic response, as assessed by 18F-FDG-PET/CT, are significant prognostic factors in patients with PCNSL.
21 Unconventional mechanisms of action and resistance to rapalogs in renal cancer
Authors: Butti Ramesh , Yang Juan , Cohn Shannon , Kapur Payal , Brugarolas James
Published: August 2024

Background

Clear cell renal cell carcinoma (ccRCC) is a prevalent cancer type in the United States driven by inactivation of the von Hippel-Lindau (VHL) gene and with frequent hyperactivation of mammalian target of rapamycin complex 1 (mTORC1). Multiple drugs have been developed to target VEGF/VEGFR2 and mTORC1 for advanced RCC treatment. Two specific mTORC1 inhibitors, temsirolimus and everolimus (known as rapalogs), are FDA-approved for the treatment of RCC but their clinical efficacy is hindered by the emergence of resistance.

Methods

To study the development of rapalog resistance in RCC, we utilized patient-derived tumorgrafts (TGs) implanted in immunocompromised mice and treated the mice with rapamycin for prolonged periods to generate resistance. Resistance was studied by transplanting resistant tumors into additional cohorts of mice and establishing primary cultures. The impact of rapamycin on tumor growth and mTORC1 activity in both tumor and stromal cells was assessed using molecular techniques. To dissect the role of the tumor microenvironment (TME), we generated genetically engineered immunocompromised recipient mice with an mTOR inhibitor resistance mutation (S2035T). Coculture experiments were performed to further explore the interplay.

Results

Prolonged rapamycin treatment resulted in the development of resistance in TGs. Notably, this occurred despite persistent inhibition of mTORC1 in tumor cells. Resistance was transient and lost with subsequent transplantation and in cell culture. Surprisingly, resistance was accompanied by mTORC1 reactivation in the TME, specifically in cancer associated fibroblasts. Introducing an mTOR resistance mutation (S2035T) into recipient mice was sufficient to cause resistance. Co-culture experiments with fibroblasts further demonstrated that rapamycin-resistant mutant fibroblasts conferred resistance in tumor cells even in the absence of mTORC1 activity.

Conclusions

This study uncovers a critical role of the TME, in mediating the response and resistance to rapalogs in RCC. Our data show that inhibition of mTORC1 in non-tumor cells is essential for the anti-tumor activity of rapalogs. These findings shed light on why mTOR mutations are rarely observed in resistant RCC patients and highlight the significance of the TME in modulating drug response and resistance. Moreover, our study emphasizes the potential of targeting TME cells as a therapeutic strategy in RCC and possibly other cancers.

DOD CDMRP Funding: yes
58 Tumor immune microenvironment determinants of response to Lenvatinib and aPD-1 blockade in clear cell renal cell carcinoma
Authors: Vuong Lynda
Published: August 2024

Background

Combinations of immune checkpoint blockade (ICB) and anti-angiogenesis tyrosine kinase inhibitors (TKI) are the mainstay of metastatic ccRCC front-line treatment, but heterogenous responses and lack of biomarkers strongly emphasizes the need to understand RCC-specific mechanisms of effective treatment-related anti-tumor immunity. Although prior work consistently highlights tumor associated macrophages (TAMs) in RCC prognosis and treatment response, TAM targeting has yet to experimentally assessed as a viable treatment sensitizer.

Methods

We therefore assessed the impact of TAM depletion using a clinical-grade colony stimulating factor receptor-1 inhibitor (CSF1Ri) on ICB or TKI sensitivity in a spontaneous mouse model of ccRCC with inducible kidney-specific Vhl, p53 and Rb1 deletions (KVpR). Mice were randomized to receive either no treatment, single agent Lenvatinib (TKI), aPD-1, or combinations with CSF1Ri. We longitudinally monitored a total of 307 tumors from 96 mice using magnetic resonance imaging (MRI) and collected responding and non-responding tumors for single cell RNA sequencing (scRNAseq) to elucidate the underlying mechanisms of response.

Results

Surprisingly we found that TAM depletion abolished aPD-1 response, with no effect on Lenvatinib response. ScRNAseq analyses suggest that CSF1R selectively depletes antigen presenting TAMs, leading to a striking loss of T cell infiltration. This suggests that TAMs, rather than dendritic cells as shown in other cancer types, may play a dominant role in eliciting T cells during aPD-1 therapy in RCC. On the other hand, Lenvatinib, which elicited a stronger T cell response than aPD-1, was associated with significant increases in cDC1 and plasma cells in responders, together suggesting that these two treatment modalities may operate through distinct antigen-presenting cells. Also unexpectedly, we found that both Lenvatinib and CSF1Ri induced severe intratumor hypoxia, which is typically associated with poor outcomes in most cancers. We identify a hypoxic niche-specific TAM subset that when abundant at baseline is predictive for poor response to ICB/TKI combos across multiple RCC trials, but post-treatment seems to capture hypoxia-mediated tumor cell death in responders in mouse and humans. We therefore wondered whether high levels of existing tumor hypoxia would exacerbate TKI-induced hypoxia leading to worst outcomes and found that in two novel RCC syngeneic models with high-baseline hypoxia, TKI indeed augments metastatic disease.

Conclusions

Our results offer mechanistic insight into the failure of CSF1R-based TAM depletion approaches in clinical trials and presents cautionary evidence against the use of hypoxia-inducing agents in tumors with high-baseline hypoxia. We show that TKI and ICB-induce distinct TME and hypoxia-related changes which require further study. More generally our study strongly emphasizes the importance of developing mechanism-guided clinical biomarkers.

DOD CDMRP Funding: yes
Racial-Ethnic Poverty Gaps in Later Life: A Role for Late Career Employment Quality?
Authors: Phillips Lora A , Rhodes Alec P
Published: August 2024

Racial-ethnic disparities in poverty are an important form of inequality in older age. Recent scholarship on racial-ethnic poverty gaps demonstrates that, beyond individual characteristics and behaviors, racialized structural factors like employment contribute to such gaps. Yet surprisingly little is known about the role of employment quality, despite observed racial-ethnic disparities in employment quality and the role of employment history in shaping later life well-being. Using data from the 2002–2018 waves of the Health and Retirement Study (HRS) and three poverty measures, we decompose the proportions of the Black-white and Hispanic-white poverty gaps among households led by 65-year-olds that are attributable to disparities in late career employment quality. We find that racial-ethnic disparities in late career employment quality account for 17–28 percent of the observed Black-white and 18–32 percent of the observed Hispanic-white poverty gaps, thus explaining a greater proportion of such gaps than many common individual or behavioral explanations. Disaggregating employment quality into its component measures, we find racial-ethnic disparities in access to employer-provided health insurance and hourly wages account for the largest proportion of racial-ethnic poverty gaps. Our findings suggest that employment quality captures important racialized dimensions of labor market context that help account for racial-ethnic inequalities in later life poverty in the United States.
Assessment of B-type natriuretic peptide as an early predictor of mortality in acutely poisoned patients with cardiotoxicities
Authors: Elgendy Shaimaa Ali , Shoeib Osama , Elgharbawy Doaa , Abo El-Noor Mona M , Kabbash Abdelmoty
Published: August 2024

Background

Cardiotoxicity is a major toxic effect induced by several types of drugs. An electrocardiogram is done routinely in cardiovascular drug exposures. Cardiac troponin I (cTnI) is the usual biomarker for diagnosing myocardial injury. B-type natriuretic peptide (BNP) is a well-established predictor of disease state in suspected heart failure.

Aim

The study aimed to assess BNP’s role as an early predictor of mortality compared with cTnI and ECG changes in acutely poisoned patients with cardiotoxicities.

Methodology

This study enrolled 70 patients with acute cardiotoxicity by drugs and toxins known to cause cardiac injury admitted to Tanta University Poison Control Center (TUPCC). Collected data included socio-demographic data, toxicological history, vital signs, ECG changes, Poison Severity Score (PSS), BNP, and cTnI serum levels.

Result

Patients were classified as survivors and non-survivors. Significantly more delay time was recorded in non-survivors. Moreover, vital signs were significantly abnormal in non-survivors. There was no statistical significance regarding the initial ECG abnormalities between survivors and non-survivors. BNP and cTnI levels were significantly higher among non-survivors. For mortality prediction, BNP had good predictive power (AUC = 0.841) with 100% sensitivity and 79.7% specificity while cTnI had an acceptable predictive power (AUC = 0.786), with 83.3% sensitivity and 78.1% specificity with insignificant difference between both biomarkers.

Conclusion

BNP and cTnI levels can predict mortality in acute cardiotoxicity compared to ECG which has no statistically significant prediction. BNP has a higher discriminatory power than cTnI for the prediction of mortality.
CPD Success With Technagogy in Health Professions: Determinants and Merits
Authors: Ong Irvin , Dino Michael Joseph , Enriquez Caroline Marian , Gotinga Trisha Joy , Esluzar Corazon , Cajayon Sharon , Buencamino Arnel , Pimentel-Tormon Fraulein , Rodriguez Angelita , Tablizo Alicia
Published: August 2024

Introduction:

Continuing professional development (CPD) has become a common strategy to address the gaps in knowledge and competencies during the pandemic. Given the drastic changes in the learning environment, this study explored “technagogy” or teaching with technology in CPD in the health professions.

Methods:

A mixed-methods study was used to ascertain the determinants and merits of CPD success from the participants' perspectives (n = 237). The quantitative data underwent structural equation modeling using partial least squares. We also thematically analyzed the qualitative responses and synthesized concurrent findings.

Results:

The structural model accounted for a 64% variance in the CPD success. Its significant direct predictors (P < .05) were instructional, curriculum, and nurturant effects. From the participants' experiences, we generated four themes as the merits of CPD: learning, teaching, knowledge, and technology scholarships. Finally, we constructed a synthesized model, the CPD Determinants and Intrinsic Assets in Learning, which can offer modest guidance in navigating CPD with technagogy.

Discussion:

The study findings emphasize the convergence and divergence points as considerations in teaching and learning with technology. It also offers valuable insights into relevant implications of technagogy in CPD, including its theory, practice, policy, and research.
Outcomes of manual arcuate keratotomy with compression sutures for high regular postkeratoplasty astigmatism
Authors: Berger Tim , Abu Dail Yaser , Seitz Berthold , Khattabi Zaynab , Flockerzi Elias , Hamon Loic , Langenbucher Achim , Daas Loay
Published: August 2024

Purpose:

To report functional and refractive outcomes of manual arcuate keratotomy (AK) with compression sutures for high regular postkeratoplasty astigmatism.

Setting:

Department of Ophthalmology, Saarland University Medical Center, Homburg/Saar, Germany.

Design:

Retrospective analysis.

Methods:

This study included 90 eyes with high regular postkeratoplasty astigmatism (>4 diopters [D]) who received AK with compression sutures between 2010 and 2022. Functional and refractive outcomes were assessed by evaluating topographic indices and by performing vector astigmatism and Fourier analysis.

Results:

At last follow-up (mean 13.7 ± 16.6 months), corrected distance visual acuity improved from 0.59 ± 0.28 to 0.34 ± 0.25 logMAR. Cylinder magnitude decreased from 9.91 ± 2.88 to 5.42 ± 3.35 D. Surface asymmetry index, irregular astigmatism index, and corneal eccentricity index were equal to preoperative values, whereas surface regularity index approached normal values at last follow-up. Fourier analysis indicated a decrease in the regular astigmatic component, whereas nonregular components (asymmetry and higher-order irregularity) remained stable. In vector astigmatism analysis, target-induced astigmatism magnitude was 9.92 ± 2.86 D and surgically induced astigmatism magnitude was 10.16 ± 4.86 D (correction index of 0.91 ± 0.48) with a difference vector of 5.42 ± 3.35 D at last follow-up. Correction of astigmatism magnitude was adequate in 40% of the eyes, undercorrected in 30%, and overcorrected in 30%. Angle of error was <|22.5 degrees| in 88% resulting in a low risk of off-axis treatment.

Conclusions:

AK with compression sutures is a simple, relatively effective, and safe surgical procedure for astigmatism reduction after keratoplasty. In case of regular astigmatism, the procedure does not increase corneal irregularities. The remaining refractive error might be further corrected by spectacles, contact lenses, or toric intraocular lens implantation (in-the-bag/add-on), thus reducing the need for repeat keratoplasty.
Long-term intrathecal infusion of low-dose morphine effectively relieves symptoms of severe restless legs syndrome/Willis–Ekbom disease without inducing opioid tolerance
Authors: Janerås Lars , Breivik Harald , Lundeland Bård , Ringstad Geir Andre , Stubhaug Audun
Published: August 2024

Restless legs syndrome/Willis–Ekbom disease (RLS/WED) causes a strong urge to move legs while resting. Restless legs syndrome/WED is an often-inherited disease occurring in 3% to 10% of adult populations, increasing with age. Severity varies from mild disturbance of sleep to painful restless legs and arms, loss of sleep, fatigue, and risk of suicide. Dopaminergic drugs relieve symptoms, but cause augmentation, ie, initially helpful but later increase the burden of symptoms. Oral gabapentinoids and opioids are often added, but opioid tolerance and adverse effects are common. With the high prevalence and incomplete help from oral drugs, significant unmet needs exist for effective therapy for severe RLS/WED. Ongoing spinal intrathecal infusion of low-dose morphine is effective, but not generally recognized, as only 12 cases have been published since 2002. We report 7 patients suffering from severe RLS/WED, who had no relief from oral dopaminergic, gabapentinoid, or opioid drugs; they all had excellent relief during ongoing spinal intrathecal infusion of morphine at only 1 to 5 μg/h, ongoing for 1 to 21 years without need of higher doses of morphine.. We suggest that morphine may be transported with the cerebrospinal fluid reaching and readjusting malfunctioning dopamine neuronal systems in the brain and spinal cord. The effects last only as long as the infusion continues. A patient with RLS/WED and persistent genital arousal disorder (PGAD) was relieved of both RLS/WED and PGAD symptoms. These case reports suggest that intrathecal infusion of low-dose morphine is an effective treatment of severe RLS.
Morphometric study of normal acetabulum on computed tomography scan in adults at a tertiary care center
Authors: Adhikari Bijay , Adhikari Pratik , Bhattarai Manoj , Pokharel Shashank , Adhikari Kapil , Gupta Mukesh Kumar
Published: August 2024

Background:

The introduction provides an in-depth understanding of the acetabulum’s anatomical structure and its significance in hip joint biomechanics. It highlights the variability in acetabular morphology among normal individuals, emphasizing the importance of establishing normal ranges for accurate diagnosis of acetabular deformities. The section also underscores the role of geographical, racial, and anatomical factors in influencing acetabular parameters, crucial for orthopedic interventions and surgical outcomes like total hip arthroplasty. Furthermore, it discusses the limitations of traditional imaging methods and the necessity of advanced techniques like computed tomography (CT) scans for precise evaluation and preoperative planning in hip surgeries.

Materials and methods:

The study adopted a cross-sectional design at a tertiary care center, enrolling 94 participants using purposive sampling for CT evaluation of acetabular morphology. Morphometric parameters including center edge angle of Wiberg, acetabular angle of Sharp, acetabular version, acetabular depth, and joint space width were measured using CT scans, while BMI categorization and statistical analyses were conducted to explore gender-based and side-based differences and correlations with BMI and age.

Results:

The study included 94 patients with ages ranging from 20 to 82 years (mean age: 49±15 years), predominantly in the 41–50 years age group. Female participants slightly outnumbered males. The mean BMI was 23.5±3.2 kg/m2, with variations across different age and BMI categories. Most morphometric parameters showed no significant differences between sexes or sides but did correlate with age and BMI, notably the center edge angle positively correlating with BMI in males.

Conclusions:

The study found correlations between morphometric parameters of the acetabulum and both age and BMI. Specifically, the center edge angle demonstrated a positive correlation with BMI in males and a weak positive correlation with age. Acetabular angle showed a negative correlation with BMI, highlighting potential implications for understanding hip joint health and conditions related to BMI changes.
Beyond the womb: pontine hemorrhage’s impact on maternal and fetal outcomes: a case report from Nepal
Authors: Yadav Jeshika , Bhusal Kiran , Thapaliya Ishwor , Bhandari Khagendra , Jha Swati S. , Khadayat Ramesh , Ghimire Asmita
Published: August 2024

Introduction:

Strokes cause 37% of cardiovascular-related deaths in women, with 40% during delivery and 50% postpartum. Pontine hemorrhage, a rare cerebrovascular disease, can occur during pregnancy and postpartum, requiring complex diagnosis and management.

Case presentation:

A 33-year-old primipara female presented with headache, vomiting, and altered consciousness at 15 weeks and 5 days gestation. She had no history of head trauma, involuntary urine passage, cough, shortness of breath, palpitation, or per vaginal bleeding. Psychiatric and neurology consultations were conducted to address her symptoms.

Discussion:

Pontine hemorrhage is a rare condition affecting pregnant women, causing mild symptoms like vertigo, ataxia, and dysphagia. Our patient underwent an initial blood investigation with imaging of her brain (CT and MRI) to establish the diagnosis of pontine hemorrhage. It is believed to be caused by hormonal fluctuations, but studies have not proven it. Stroke during pregnancy is rare, with only 10% occurring during the antepartum period. Diagnosis requires a multidisciplinary approach with history, clinical examination, and radiological imaging like CT and MRI. Treatment involved antiplatelet therapy, which ensured favorable maternal and fetal outcomes for our patient.

Conclusion:

In summary, this case demonstrates the rare incidence of pontine hemorrhage during pregnancy. It highlights the diagnostic challenges and successful treatment with antiplatelet therapy, emphasizing the necessity for a comprehensive approach to ensure favorable maternal and fetal outcomes.
Redesigning Competency Assessment
Authors: Kauffman Sharon R. , Prosdocimo Brittni
Published: August 2024

A train-the-trainer model of clinical education was redesigned in one National Cancer Institute-designated cancer center to introduce a blended model of learning. Program outcomes were for oncology nurse trainers to affirm program engagement; successfully complete a new, web-based specialized oncology curriculum, with 80% accuracy; complete one live 8-hour training event; acquire competency validation of required skills, with 100% accuracy; and communicate one strategy for training staff in their respective clinical settings.
SARS-CoV-2 infection vs. vaccination during pregnancy: the placenta leads the way
Authors: Pérez-Latorre Leire , Ramilo Octavio
Published: August 2024

Purpose of review

To understand the characteristics and determinants of transplacental antibody transfer against SARS-CoV-2 and to compare the differences between SARS-CoV-2 infection and vaccination.

Recent findings

The need for information during the COVID-19 pandemic and the exclusion of pregnant women from randomized clinical trials have led to a vast amount of clinical data primarily based on observational studies with diverse design and sample analyses that yield variable results. This review aims to critically and comprehensively integrate the relevant knowledge related to transplacental transfer of antibodies against SARS-CoV-2, emphasizing the differences between infection and vaccination.

Passive immunization is key to conferring protection to the infant during their first months of life. Understanding the mechanisms of transplacental antibody transfer during SARS-CoV-2 infection and vaccination, and their associated protection will allow optimizing the implementation of well tolerated and effective preventive strategies for both pregnant women and infants.
Association of a modified uterine manipulation method with improved recovery after laparoscopic radical hysterectomy: a retrospective cohort study
Authors: Sun Yu-Shu , Bao Ling-Jie , Xiao Yin-Ping , Chang Kai-Kai , Du Yan , Yi Xiao-Fang
Published: August 2024

Objective:

Uterine manipulation with an intrauterine device is the surgical method used for early-stage cervical cancer. In this study, we explored a modified uterine manipulation method without the using an intrauterine device during laparoscopic radical hysterectomy (LRH) of patients with early-stage cervical cancer.

Methods:

We retrospectively collected data of patients with early-stage cervical cancer who underwent LRH between January 2015 and June 2019. The clinicopathological characteristics and surgical outcomes of patients treated with the new method (Group S) were compared to those of patients treated with the conventional uterine manipulator (Group M).

Results:

A total of 100 patients were included in the study, with 54 patients in Group M and 46 patients in Group S. No significant difference was observed in clinicopathological characteristics between the two groups. Compared to Group M, Group S had a significantly shorter duration of postoperative fever (0 vs. 3 days, P = 0.003) and a shorter duration of secondary antibiotic use (4 vs. 5 days, P = 0.035). Additionally, the cost of antibiotics was lower in Group S ($95.45 vs. $135.22, P = 0.032). The modified uterine manipulation method was associated with a lower risk of postoperative fever (P = 0.007). No difference was found in the 2-year disease-free survival (P = 0.409) and 2-year overall survival between the two groups (P = 0.718).

Conclusions:

The modified uterine suture suspension method is easy to perform, enhances postoperative recovery, and could be a viable option for patients with early-stage cervical cancer.
Secreted chemokines and transcriptomic analyses reveal diverse inflammatory and degenerative processes in the intervertebral disc of the STZ-HFD mouse model of Type 2 diabetes
Authors: Gonzalez Christian E. , Vaidya Rachana S. , Clayton Sade W. , Tang Simon Y.
Published: August 2024

The chronic inflammation resultant from type 2 diabetes (T2D) is also associated with spinal pathologies, including intervertebral disc (IVD) degeneration and chronic neck and back pain. Although confounding factors, such as increased weight gain in obesity, studies have shown that even after adjusting age, body mass index, and genetics (e.g. twins), patients with T2D suffer from disproportionately more IVD degeneration and back pain. We hypothesize that chronic T2D fosters a proinflammatory microenvironment within the IVD that promotes degeneration and disrupts disc homeostasis. To test this hypothesis, we evaluated two commonly used mouse models of T2D – the leptin-receptor deficient mouse (db/db) and the chronic high-fat diet in mice with impaired beta-cell function (STZ-HFD). STZ-HFD IVDs were more degenerated and showed differential expression of chemokines from the db/db models. Moreover, the RNAseq analysis revealed vast transcriptional dysregulation of many pathways in the STZ-HFD but not in the db/db tissues. Taken together, the STZ-HFD may better recapitulates the complexities of the chronic inflammatory processes in the IVD during T2D.
Restoration of striatal neuroprotective pathways by kinase inhibitor treatment of Parkinsons linked-LRRK2 mutant mice
Authors: Jaimon Ebsy , Lin Yu-En , Tonelli Francesca , Antico Odetta , Alessi Dario R , Pfeffer Suzanne R.
Published: August 2024

Parkinsons disease-associated, activating mutations in Leucine Rich Repeat Kinase 2 (LRRK2) block primary cilia formation in cholinergic and parvalbumin interneurons and astrocytes in the striatum, decreasing the production of GDNF and NRTN neuroprotective factors that normally support dopaminergic neuron viability. We show here that 3 month-dietary administration of the MLi-2 LRRK2 kinase inhibitor restores primary cilia and the Hedgehog-responsive production of neuroprotective GDNF and NRTN by these neurons; cilia are also restored on cholinergic neurons of the pedunculopontine nucleus. Importantly, we detect recovery of striatal dopaminergic processes and decreased stress-triggered Hedgehog signaling by nigral dopaminergic neurons. Thus, pathogenic LRRK2-driven cilia loss is reversible in post-mitotic neurons and astrocytes, which suggests that early administration of specific LRRK2 inhibitors may have significant therapeutic benefit for patients in the future.
Gene editing of the E3 ligasePIRE1fine-tunes ROS production for enhanced bacterial disease resistance in tomato
Authors: Castro Bardo , Baik Suji , Tran Megann , Zhu Jie , Li Tianrun , Tang Andrea , Aoun Nathalie , Blundell Alison C , Gomez Michael , Zhang Elaine , Cho Myeong-Je , Lowe-Power Tiffany , Siddique Shahid , Staskawicz Brian , Coaker Gitta
Published: August 2024

Reactive oxygen species (ROS) accumulation is required for effective plant defense. Accumulation of the Arabidopsis NADPH oxidase RBOHD is regulated by phosphorylation of a conserved C-terminal residue (T912) leading to ubiquitination by the RING E3 ligase PIRE. ArabidopsisPIREknockouts exhibit enhanced ROS production and resistance to the foliar pathogenPseudomonas syringae. Here, we identified 170PIREhomologs, which emerged in Tracheophytes and expanded in Angiosperms. We investigated the role ofSolanum lycopersicum(tomato) PIRE homologs in regulating ROS production, RBOH stability, and disease resistance. Mutational analyses of residues corresponding to T912 in the tomato RBOHD ortholog, SlRBOHB, affected protein accumulation and ROS production in aPIRE-dependent manner. Using CRISPR-cas9, we generated mutants in twoS. lycopersicum PIREhomologs (SlPIRE).SlPIRE1edited lines (Slpire1) in the tomato cultivar M82 displayed enhanced ROS production upon treatment with flg22, an immunogenic epitope of flagellin. Furthermore, Slpire1exhibited decreased disease symptoms and bacterial accumulation when inoculated with foliar bacterial pathogensPseudomonas syringaeandXanthomonas campestris. However,Slpire1exhibited similar levels of colonization as wild type upon inoculation with diverse soilborne pathogens. These results indicate that phosphorylation and ubiquitination crosstalk regulate RBOHs in multiple plant species, andPIREis a promising target for foliar disease control. This study also highlights the pathogen-specific role ofPIRE, indicating its potential for targeted manipulation to enhance foliar disease resistance without affecting root-associated interactions, positioningPIREas a promising target for improving overall plant health.
snRNA-seq analysis of the mossPhyscomitrium patensreveals a conserved cytokinin-ESR module promoting pluripotent stem cell identity
Authors: Hata Yuki , Hetherington Nicola , Battenberg Kai , Hirota Atsuko , Minoda Aki , Hayashi Makoto , Kyozuka Junko
Published: August 2024

The shoot apical meristem (SAM), which contains pluripotent stem cells, serves as the source of the entire shoot system in land plants. To uncover the mechanisms underlying SAM development and its orign, we employed single-nucleus RNA sequencing technology inPhyscomitrium patens, which contains a single stem cell known as the gametophore apical cell. We identified distinct cell clusters representing major cell types of theP. patensgametophyte, including the gametophore apical cells. We revealed dynamic gene expression changes during cell fate progression in the gametophore apical cell and found upregulation of cytokinin biosynthesis genes in this cell. We also identifiedENHANCER OF SHOOT REGENERATION 1(ESR1) orthologs as key regulators of gametophore apical cells downstream of cytokinin. Given that ESRs promote SAM formation under cytokinin in angiosperms, we propose that the cytokinin-ESR module represents a conserved mechanism promoting stem cell identity that evolved in the common ancestor of land plants.
The assembly of Y chromosome reveals amplification of genes regulating male fertility inBactrocera dorsalis
Authors: Wu Shuangxiong , Wu Jiahong , Lei Quan , He Donghai , Jiang Xinrui , Ye Chao , Wei Dong , Wang Jinjun , Xu Luohao , Jiang Hongbo
Published: August 2024

BACKGROUND

The oriental fruit flyBactrocera dorsalisis an invasive pest causing considerable ecological and economic damage worldwide. The Y chromosome is an important target for the sterile insect technique (SIT) but its sequence and structure has been poorly explored.

RESULTS

We assembled the genome ofB. dorsalisat the chromosome level with a total size of 562.6 Mb. The assembly includes a ∼7.6 Mb Y chromosome sequence, being the first reported Y chromosome in Tephritidae. The assembled Y chromosome is highly repetitive, and contains 61 genes, including 9 multi-copy genes. We surprisingly found that the M factor (MoY) in Tephritidae has multiple copies, verified by our droplet digital PCR (ddPCR) analysis. Besides, we identified 32 copies ofgyf-like on the Y chromosome(gyfY) that were highly expressed in testis. RNAi targeting thegyfYresulted in depletion of live sperms, suggesting that the amplification ofgyfYis essential for male fertility, which facilitated the understanding of high fecundity of this invasive insect.

CONCLUSION

We reported firstly the Y chromosome ofBactrocera dorsalis. Our results will also provide target genes for CRISPR/Cas9 based SIT, leading to the development of novel control strategies against tephritid flies.
Distinct microbial communities are linked to organic matter properties in millimetre-sized soil aggregates
Authors: Simon Eva , Guseva Ksenia , Darcy Sean , Alteio Lauren , Pjevac Petra , Schmidt Hannes , Jenab Kian , Ranits Christian , Kaiser Christina
Published: August 2024

Soils provide essential ecosystem services and represent the most diverse habitat on Earth. It has been suggested that the presence of various physico-chemically heterogenous microhabitats supports the enormous diversity of microbial communities in soil. However, little is known about the relationship between microbial communities and their immediate environment at the micro- to millimetre-scale. In this study, we examined whether bacteria, archaea, and fungi organise into distinct communities in individual 2-millimetre-sized soil aggregates and compared them to communities of homogenized bulk soil samples. Furthermore, we investigated their relationship to their local environment by concomitantly determining microbial community structure and physico-chemical properties from the same individual aggregates. Aggregate-communities displayed exceptionally high beta-diversity, with 3-4 aggregates collectively capturing more diversity than their homogenized parent soil core. Up to 20-30% of ASVs (particularly rare ones) were unique to individual aggregates selected within a few centimetres. Aggregates and bulk soil samples showed partly different dominant phyla, indicating that taxa that are potentially driving biogeochemical processes at the small scale may not be recognized when analysing larger soil volumes. Microbial community composition and richness of individual aggregates were closely related to aggregate-specific carbon and nitrogen content, carbon stable-isotope composition, and soil moisture, indicating that aggregates provide a stable environment for sufficient time to allow co-development of communities and their environment. We conclude that the soil microbiome is a metacommunity of variable subcommunities. Our study highlights the necessity to study small, spatially coherent soil samples to better understand controls of community structure and community-mediated processes in soils.
A Library of Custom PEG-Lipids reveals a Double-PEG-Lipid with Drastically Enhanced Paclitaxel Solubility and Human Cancer Cell Cytotoxicity when used in Fluid Micellar Nanoparticles
Authors: Ghasemizadeh Aria , Wan Lili , Hirose Aiko , Diep Jacqueline , Ewert Kai K. , Safinya Cyrus R.
Published: August 2024

Paclitaxel (PTX) is one of the most widely utilized chemotherapeutics globally. However, the extremely poor water solubility of paclitaxel necessitates a mechanism of delivery within blood. Fluid lipid PTX nanocarriers (lipids in the chain-melted state) show promise as PTX delivery vectors, but remain limited by their solubility of PTX within the membrane. To improve pharmacokinetics, membrane surfaces are typically coated with polyethylene glycol (PEG). Recent work has demonstrated the generation of a population of micelles within fluid lipid formulations containing a 2kDa PEG-lipid at a 10 mol% ratio. Driven by the positive curvature of the PEG-lipid (i.e. area of head group > area of tails), micelle-containing formulations were found to exhibit significantly higher uptake in cancer cells, cytotoxicity, andin vivoantitumor efficacy compared to formulations containing solely liposomes. Here, we describe the custom synthesis of a library of high-curvature micelle-inducing PEG-lipids and examine the effects of PEG chain length, chain branching (single- or double-PEG-lipid), and cationic charge on PTX solubility and cytotoxicity. We examined PEG-lipids at standard (10 mol%) and high (100-x mol%, where x=PTX mol%) formulation ratios. Remarkably, all formulations containing the synthesized high-curvature PEG-lipids had improved PTX solubility over unPEGylated formulations and commercially available DOPE-5k. The highest PTX solubility was found within the 100–xptxmol% PEG-lipid micellar formulations, with particles made from 2k2(two PEG2k chains) encapsulating 13 mol% PTX for up to 24 h. The pancreatic cancer cell line PC3 exhibited higher sensitivity to formulations containing PEG-lipid at 100–xptxmol%, the most potent of which being formulations made from 2k2(IC50 = 14 nM). The work presented here suggests formulations employing high-curvature PEG-lipids, particularly the double-PEG-lipid 2k2, hold great potential as next-generation PTX delivery systems owing to their high PTX solubility, enhanced cell cytotoxicity, and ability for precision targeting by affixation of ligands to the PEG molecules.
The AP2/ERF transcription factorHcERF5confers drought tolerance via ABA-mediated signaling in kenaf
Authors: Luo Dengjie , Mubeen Samavia , Rehman Muzammal , Cao Shan , Wang Caijin , Yue Jiao , Pan Jiao , Jin Gang , Li Ru , Chen Tao , Chen Peng
Published: August 2024

The APETALA2/ethylene response factor (AP2/ERFs) are pivotal in regulating abiotic stress responses in plants. However, the specific role of ERFs in kenaf’s response to drought stress remains unclear. In this study, a transcription factorsHcERF5was isolated from kenaf and its role in drought stress tolerance was analyzed. HcERF5 was found to localize in both nucleus and cytoplasm and could be significantly induced by polyethylene glycol-6000 (PEG-6000) and abscisic acid (ABA) in kenaf seedlings. In transgenicArabidopsisexpressing theHcERF5promoter-driven β-glucuronidase (GUS), strong GUS activity was observed in roots, stems, and leaves. Overexpression ofHcERF5inArabidopsisenhanced seed germination rates under drought or ABA stress and improved drought tolerance in seedlings by increasing antioxidant enzyme activities, whereasaterf5knockout lines exhibited the opposite trend. Additionally,HcERF5overexpressingArabidopsisshowed significantly increased drought tolerance and reduced sensitivity to ABA. Furthermore, virus-induced gene silencing (VIGS) ofHcERF5in kenaf reduced drought tolerance, as evidenced by decreased antioxidant enzyme activity, increased stomatal aperture, and elevated levels of malondialdehyde (MDA), reactive oxygen species (ROS), and proline under drought stress. RNA-seq analysis further revealed thatHcERF5directly regulated ABA signaling pathway. Yeast-two-hybrid (Y2H) assays revealed 29 proteins that interact with HcERF5. Among them, the expression of downstream drought stress-related genesHcPRK,HcRD22,HcMAP2,HcCAB,HcCS, andHcCCoAOMT3were significantly reduced inHcERF5-silenced plants. Overall, this study highlights the significant potential ofHcERF5 in enhancing drought tolerance in kenaf.

Highlight

HcERF5overexpression enhanced drought tolerance in kenaf, however silencing increased drought sensitivity.

HcERF5regulates ABA synthesis and increases kenaf stomatal conductance and density under drought stress.

HcERF5regulates plant hormone signal transduction, MAPK signalling, and phenylpropanoid biosynthesis in kenaf under drought stress.

Protein interaction revealed HcERF5 interacts with six stress-response genes.
Optogenetic control of phosphate-responsive genes using single component fusion proteins inSaccharomyces cerevisiae
Authors: Cleere Matthew M. , Gardner Kevin H.
Published: August 2024

Blue light illumination can be detected by Light-Oxygen-Voltage (LOV) photosensing proteins and translated into a range of biochemical responses, facilitating the generation of novel optogenetic tools to control cellular function. Here, we develop new variants of our previously described VP-EL222 light-dependent transcription factor and apply them to study the phosphate-responsive signaling (PHO) pathway in the budding yeastSaccharomyces cerevisiae, exemplifying the utilities of these new tools. Focusing first on the VP-EL222 protein itself, we quantified the tunability of gene expression as a function of light intensity and duration, and demonstrated that this system can tolerate the addition of substantially larger effector domains without impacting function. We further demonstrated the utility of several EL222-driven transcriptional controllers in both plasmid and genomic settings, using thePHO5andPHO84promoters in their native chromosomal contexts as examples. These studies highlight the utility of light-controlled gene activation using EL222 tethered to either artificial transcription domains or yeast activator proteins (Pho4). Similarly, we demonstrate the ability to optogenetically repress gene expression with EL222 fused to the yeast Ume6 protein. We finally investigated the effects of moving EL222 recruitment sites to different locations within thePHO5andPHO84promoters, as well as determining how this artificial light-controlled regulation could be integrated with the native controls dependent on inorganic phosphate (Pi) availability. Taken together, our work expands the applicability of these versatile optogenetic tools in the types of functionalities they can deliver and biological questions that can be probed.
Respiratory Syncytial Virus (RSV) optimizes the translational landscape during infection
Authors: Kerkhofs Kyra , Guydosh Nicholas R. , Bayfield Mark A.
Published: August 2024

Viral infection often triggers eukaryotic initiator factor 2α (eIF2α) phosphorylation, leading to global 5’-cap-dependent translation inhibition. RSV encodes messenger RNAs (mRNAs) mimicking 5’-cap structures of host mRNAs and thus inhibition of cap-dependent translation initiation would likely also reduce viral translation. We confirmed that RSV limits widespread translation initiation inhibition and unexpectedly found that the fraction of ribosomes within polysomes increases during infection, indicating higher ribosome loading on mRNAs during infection. We found that AU-rich host transcripts that are less efficiently translated under normal conditions become more efficient at recruiting ribosomes, similar to RSV transcripts. Viral transcripts are transcribed in cytoplasmic inclusion bodies, where the viral AU-rich binding protein M2-1 has been shown to bind viral transcripts and shuttle them into the cytoplasm. We further demonstrated that M2-1 is found on polysomes, and that M2-1 might deliver host AU-rich transcripts for translation.

Importance

Viruses strongly rely on the host’s translational machinery to produce viral proteins required for replication. However, it is unknown how viruses that do not globally inhibit cap-dependent translation compete with abundant host transcripts for ribosomes. In this study, we found that respiratory syncytial virus (RSV) infection results in redistribution of 80S monosomes into the polysomes. High-throughput sequencing of translating transcripts revealed that low translation efficiency transcripts become more efficient at ribosome recruitment which are virus-resembling AU-rich host transcripts. Finally, we also uncover that AU-rich RNA binding protein RSV-M2-1 interacts with polysomes through contacts to mRNA. These findings revealed that RSV optimizes the translational landscape rather than inhibiting host translation.
Concerted modulation of spontaneous behavior and time-integrated whole-brain neuronal activity by serotonin receptors
Authors: Friedmann Drew , Gonzalez Xavier , Moses Ashley , Watts Tanner , Degleris Anthony , Ticea Nicole , Song Jun H , Datta Sandeep Robert , Linderman Scott W , Luo Liqun
Published: August 2024

Serotonin neurons from the raphe nuclei project across the entire brain and modulate diverse physiology and behavior by acting on over a dozen receptors. Here, we took a step towards dissecting this complex process by examining the effects of agonists and antagonists of four widely expressed serotonin receptors (2A, 2C, 1A, and 1B) on spontaneous mouse behavior, which we related to time-integrated whole-brain neuronal activity as assessed by the expression of Fos, a canonical immediate-early gene product. Low-dimensional representations of behavioral and Fos map data revealed the dominant factors of variation in each domain, captured predictable differences across drug groups, and enabled predictions of behavioral changes following perturbations in Fos maps and vice versa. Our study provides a rich resource describing the effects of manipulating serotonin receptors on animal behavior and whole-brain integrated neuronal activity. It also establishes an experimental and analysis paradigm for interrogating the relationship between behavior and neuronal activity across different time scales.
Unraveling the Molecular mechanism of Polysaccharide Lyases for Efficient Alginate Degradation
Authors: Rivas-Fernández José Pablo , Vuillemin Marlene , Pilgaard Bo , J. Klau Leesa , Fredslund Folmer , Lund-Hanssen Charlotte , H. Welner Ditte , S. Meyer Anne , Morth J. Preben , Meilleur Flora , L. Aachmann Finn , Rovira Carme , Wilkens Casper
Published: August 2024

Alginate lyases (ALs) are essential for breaking down brown macroalgae alginates, widely used naturally-occurring polysaccharides. Their molecular mechanisms remain challenging due to the lack of catalytically competent Michaelis-Menten complex structures. We here provide structural snap-shots and dissect the mechanism of mannuronan-specific ALs from family 7 polysaccharide lyases (PL7), employing time-resolved NMR, X-ray, neutron crystallography, and QM/MM simulations. We reveal the protonation state of critical active site residues, enabling atomic-level analysis of the reaction coordinate. Our approach reveals an endolytic and asynchronous syn β-elimination reaction, with Tyr serving as both Brønsted base and acid, involving a carbanion-type of transition state. This study not only reconciles previous structural and kinetic discrepancies, but also establishes a comprehensive PL reaction mechanism applicable across lyase families, which can guide the engineering of ALs for tailored alginate oligosaccharide production.