bluuebunny/crossref_metadata_2025 huggingface dataset:
(72) THE HABITS OF EUROPEAN UROLOGISTS IN THE FIELD OF CRYOPRESERVATION BEFORE UROLOGICAL CANCERS TREATMENT
Authors: Degraeve Dr Amandine
Published: June 2024

Objectives

Treatments against urogenital cancers frequently have fertility side-effects. The strategy to preserve fertility after oncologic treatments is still a matter of debate with a lack of evidence and international guidelines. The aim of this study is to investigate fertility preservation practices before urogenital cancers treatments and to compare national habits.

Methods

An online anonymous survey was submitted from January to June 2021 to 6 European urological societies. The 31-items questionnaire included questions about demography, habits of evaluation and management of fertility preservation in case of urogenital cancers treatments.

Results

228 urologists from six urological societies in five different countries (Belgium, The Netherlands, Luxembourg, France, Finland) filled out the survey.

Three quarter (74%; n = 166) usually propose a cryopreservation before orchidectomy. In case of oligo/azoo-spermia, the technique performed for the sperm extraction during orchidectomy varies among the sample: 70.5% (n=160) of the responders do not perform a Testicular Sperm Extraction (TESE) nor a Percutaneous Epididymal Sperm Aspiration (PESA).

The cryopreservation for prostate cancer treatments is never proposed in 48.17% (n = 105) of responders but conversely it is always proposed in 5.05% (n = 11).

The cryopreservation before bladder cancer treatments is not commonly proposed (67.5%, n = 154).

Conclusions

Our study showed variable country specific tendencies in terms of fertility preservation in the period of treatment of urological cancers. These differences seem to be related to national guidelines and recommendations. International guidelines are urgently needed in the field of fertility for urological cancer patients. It is of utmost importance to take a clear stance on the potential importance of the semen preservation before orchidectomy, TESE and PESA in the tumor-bearing testis during orchidectomy in case of azoospermia and semen preservation discussion for young patients.

Conflicts of Interest

No conflict of interest.
(183) A PILOT STUDY OF POST-VESTIBULECTOMY OPIOID USE FOLLOWING PRE-OPERATIVE VESTIBULAR LOW INTENSITY SHOCKWAVE THERAPY
Authors: Goldstein Sue W , Goldstein Dr Irwin
Published: June 2024

Objectives

Low intensity shockwave therapy (LiWST) is FDA-cleared for pain amelioration. We serendipitously observed that one patient trying to avoid vestibulectomy surgery for neuroproliferative vestibulodynia (NPV) by undergoing multiple vestibular LiWST treatments used very few narcotics post-vestibulectomy. As a result, we studied whether using pre-operative LiSWT would decrease post-operative opioid use in patients undergoing complete vestibulectomy for NPV.

Methods

Post-op opioid use among women who underwent complete vestibulectomy at our facility January 1, 2019 to June 30, 2021 was assessed based on patient’s recall of total oxycodone/acetaminophen tablets required within 30 days post-vestibulectomy. When possible, patients confirmed use by counting tablets remaining in the pill container. Women scheduled for vestibulectomy were included in the LiSWT study group if they agreed to undergo 3 vestibular LiSWT treatments immediately pre-operatively. Using the Urogold 100™MTS, LiSWT involved delivery of 1800 shocks to the vestibule per session, energy flux density 0.05-0.1 mJ/mm2, 3 Hz, membrane pressure 3. We compared post-op opioid use by women who did and did not undergo pre-operative LiSWT.

Results

Forty-five women (mean age 28 years) underwent complete vestibulectomy during the 30-month period. Twenty women (mean age 27 years) underwent pre-op LiSWT and 25 (mean age 29 years) did not. Post-op opioid use was obtained from all but 6 in the non-LiSWT group. Mean post-op opioid use was 22 +/- 10 (range 1-26) tablets in patients who underwent vestibular LISWT treatments; opioid side effects were restricted to constipation, sleepiness, and nausea. Mean post-operative opioid use was 47 +/- 13 (range 25-90) tablets in those without vestibular LiSWT; opioid side effects included constipation, nausea, fainting, drowsiness, fatigue, and blurred vision.

Conclusions

In this pilot study, pre-operative low intensity shockwave therapy decreased post-operative opioid use after complete vestibulectomy with vaginal advancement flap. While encouraging, more research is needed.

Conflicts of Interest

No conflicts of interest.
(185) SAFETY AND PRELIMINARY EFFICACY OF PHASE 1 TRIAL TARGETING HPV16 INFECTION AND CERVICAL LESIONS WITH INTERFERON-INDUCING VIRAL VECTOR DELNS/E6E7: A PLACEBO-CONTROLLED STUDY WITH 28 PATIENTS
Authors: Bayerle-Eder M M D , Fuchs C , Muster T , Nicolodi C , Groiss F , Wolzt M
Published: June 2024

Objectives

Human papillomavirus type 16 (HPV16) is a major causative agent of cervical cancer, necessitating the development of innovative therapeutic strategies. This Phase 1 clinical trial aimed to evaluate the safety and preliminary efficacy of a novel intervention against persistent HPV16 infection and associated cervical lesions, utilizing an interferon-inducing viral vector expressing tumor antigens (delNS/E6E7), in a placebo-controlled study involving 28 patients.

Methods

In this placebo-controlled Phase 1 trial, 28 participants with HPV16 infection and cervical lesions were randomly assigned in a 3:1 ratio to receive the interferon-inducing viral vector delNS/E6E7 or a placebo over a 12-week period. Safety assessments were conducted regularly throughout the trial, and efficacy was evaluated by monitoring changes in HPV16 infection and cervical lesion status.

Results

The intervention group, which received delNS/E6E7, exhibited a favorable safety profile, with only mild and moderate transient adverse events reported. No serious adverse events related to the treatment were observed. Early indications of efficacy, such as HPV 16 clearance and regression or stabilization of cervical lesions were observed. The 12 month period of the follow-up study is still ongoing.

Conclusions

Conclusion: In this placebo-controlled Phase 1 trial, the interferon-inducing viral vector delNS/E6E7 exhibited a favorable safety profile and early indications of efficacy for the treatment of HPV16 infection and associated cervical lesions. These findings emphasize the potential of this innovative treatment approach for HPV16-related cervical health issues and underscore the need for further investigation in larger, randomized, controlled trials.

Conflicts of Interest

F. Groiss, T. Muster and C. Nicolodi are working for the company, which is producing the immune therapy (Blue Sky GmbH, Vienna, Austria).

M. Bayerle-Eder is consultant of Blue Sky GmbH.
(146) ASSESSING SEXUAL IMPULSE DEVELOPMENT SKILLS USING THE TCS-A IN A GROUP OF ADOLESCENTS: RESULTS AND IMPLICATIONS FOR DESIGN AND EDUCATIONAL INTERVENTION
Authors: Buono Dr Gabriele , Chiapparino Dr Marco , Denegri Dr Martina , Caprile Cristina , Pastorino Sabrina , Sferrazzo Dr Francesco
Published: June 2024

Objectives

The objective is to know the current condition of adolescents in the province in order to build a project of affectivity and sexuality education according to a bottom-up approach, taking into account emerging needs. However, due to the numerical limitation, it is intended to reach a more representative sample in collaboration with the Regional School Office and the Liguria Health Authority.

Methods

The TCS-A test was administered anonymously to 40 two-year adolescents (first and second classes) of a technical high school in Imperia, including 27 malesand 13 females between the ages of 14 and 19. Data were analyzed using Jamovi software. The test assesses cognitive, relational, sexual, and self-identity-building developmental skills.

Results

The research found that the area of sexual drive development competence is the most deficient compared to the other areas (T=29.9): in particular, it is statistically significant that the mean of the overall factor decreases with increasing age (p=0.032; r=-0.296), consistent with class (p=0.026; r=-0.31). For the area of sexual drives, it is statistically significant that as age increases, the following decreases: coping with sexual drives (p=0.04; r=-0.28), pride related to sexual development (p=0.017, r=-0.335), guilt-free sexual experience (p=0.04; r=-0.281), and feeling of difficulty in having a romantic relationship (p=0.084, r=-0.281).

Conclusions

Respect to the other developmental competencies, the scores are within the average (±1sd), except for the personal identity construction competence (39.1), particularly the area of life satisfaction (34.5). Respect to the open-ended questions, it emerges that those who would like to engage in sexuality education, define it in a more complex way, and those who would not like to participate in a project links sexuality to the physical dimension alone. The needs could be: scientifically based knowledge about sexuality, inclusive sexuality education, and sexual and general well-being.

Conflicts of Interest

The authors declare no conflict of interest.
(411) PROJECT BRIDGE: LINKING AT-RISK INDIVIDUALS TO A SUITABLE PREVENTATIVE CARE BASED ON RISK LEVEL
Authors: Jozefík Dominik , Ballester-Arnal Prof Rafael , Briken Prof Peer , Klapilová Kateřina , Metenko Prof Jozef , Vaaranen-Valkonen Dr Nina , Rahm Christoffer
Published: June 2024

Objectives

Projekt BRIDGE is an international two-year research project, funded by the European Commission under the Internal Security Fund, aimed at developing and evaluating two anonymous online interventions for individuals with a sexual interest in minors searching and using CSAM in SurfaceWeb and DarkWeb that are tailored to participant's risk level of sexually offending.

Methods

Two interventions tested are: (1) MiBridge – a three-week, anonymous, online Motivational Interviewing intervention aiming to motivate high-risk participants to admit to treatment face-to-face as identified patients, and (2) ReDirection – a five-week, anonymous online cognitive behavioural self-help program intervention aiming to reduce sexual urges and behaviours regarding children in the group with low/medium risk participants. Within the project, MiBridge and ReDirection are translated into several European languages and tested for feasibility in the randomised pilot trial. Based on the results, revised versions are evaluated for efficacy, safety and appreciation in two pre-registered, multicenter, randomised and waitlist-controlled trials. Inclusion criteria for all trials: English-, Swedish-, Spanish-, Finish-, Czech-, Slovak- or German-speaking; 18 years or older; with sexual urges directed towards children and past six months use of CSAM.

Results

The project started from January 2023 till December 2024. Recruitment of participants for the pilot trial ran from 30.06.2023 to 12.09.2023, and recruitment of participants for the randomised controlled trial is running from 14.09.2023 to 21.04.2024. During the two months of piloting, 357 people expressed interest in joining the project, of which 152 people booked an initial interview, and 11 people subsequently graduated from the project (5 ReDirection; 6 MiBridge).

Conclusions

The project represents a pioneering effort to address the sensitive and challenging issue of CSAM innovatively by implementing intervention strategies to make treatment available to those at risk of committing a sexually motivated offence. Preliminary results of the recruitment strategies and the efficacy of the interventions of the randomised control trial will be presented as part of the conference presentation.

Conflicts of Interest

No conflicts of interest.
Unique presentation of acute neuro-Behçet’s disease involving a cytotoxic oedema core surrounded by vasogenic oedema
Authors: Yamashita Shohei , Fujimori Daiki , Igari Shigemoto , Yamamoto Yusuke , Mizuuchi Takahiro , Mori Hiroaki , Hayashi Haeru , Tahara Koichiro , Sawada Tetsuji
Published: June 2024

A 53-year-old woman with recurrent stomatitis, genital ulcers, and folliculitis was admitted to Tokyo Medical University Hospital after experiencing visual disturbances for the past 2 weeks and a nonthrobbing headache for the past 3 days. She had also developed numbness in her left extremities. An ophthalmological examination revealed inflammatory changes in the eye. Cerebrospinal fluid analysis showed increased cell counts, protein, and interleukin-6 levels. Brain magnetic resonance imaging revealed multiple high signal intensities on T2-weighted/fluid-attenuated inversion recovery images of the pons and occipital and parietal lobes. The T2-weighted/fluid-attenuated inversion recovery high-signal-intensity lesion in the pons was hyperintense on diffusion-weighted imaging and hypointense on apparent diffusion coefficient mapping, suggesting cytotoxic oedema. Another high-signal-intensity lesion on T2-weighted/fluid-attenuated inversion recovery was isointense to hyperintense on diffusion-weighted imaging and hyperintense on apparent diffusion coefficient, indicating vasogenic oedema. The vasogenic oedema in the left occipital lobe contained a small core that was hyperintense on diffusion-weighted imaging and hypointense on apparent diffusion coefficient, suggesting cytotoxic oedema. The patient was diagnosed with acute neuro-Behçet’s disease and responded well to high-dose glucocorticoid and colchicine treatment. The present report emphasises that patients with acute neuro-Behçet’s disease may present with cytotoxic oedema in the pons and cerebral spheres. Further reports of similar cases would contribute to a better understanding of the role of cytotoxic oedema in the pathophysiology of neuro-Behçet’s disease and help elucidate the mechanisms underlying a unique presentation characterised by a central cytotoxic oedema core within vasogenic oedema.
Rmg10, a Novel Wheat Blast Resistance Gene Derived from Aegilops tauschii
Authors: Yoshioka Motohiro , Kishii Masahiro , Singh Pawan Kumar , Inoue Yoshihiro , Vy Trinh Thi Phuong , Tosa Yukio , Asuke Soichiro
Published: June 2024

Wheat blast, caused by Pyricularia oryzae (syn. Magnaporthe oryzae) pathotype Triticum (MoT), is a devastating disease that can result in up to 100% yield loss in affected fields. To find new resistance genes against wheat blast, we screened 199 accessions of Aegilops tauschii, the D genome progenitor of common wheat ( Triticum aestivum), by seedling inoculation assays with Brazilian MoT isolate Br48 and found 14 resistant accessions. A synthetic hexaploid wheat line (Ldn/KU-2097) derived from a cross between the T. turgidum ‘Langdon’ (Ldn) and resistant A. tauschii accession KU-2097 exhibited resistance in seedlings and spikes against Br48. In an F2 population derived from ‘Chinese Spring’ × Ldn/KU-2097, resistant and susceptible individuals segregated in a 3:1 ratio, suggesting that the resistance from KU-2097 is controlled by a single dominant gene. We designated this gene Rmg10. Genetic mapping using an F2:3 population from the same cross mapped the RMG10 locus to the short arm of chromosome 2D. Rmg10 was ineffective against Bangladesh isolates but effective against Brazilian isolates. Field tests in Bolivia showed increased spike resistance in a synthetic octaploid wheat line produced from a cross between common wheat cultivar ‘Gladius’ and KU-2097. These results suggest that Rmg10 would be beneficial in farmers’ fields in South America.
A Severe and Prolonged Case of Ocular Monkeypox Without Systemic Manifestations
Authors: Shan Kevin , Montalvo-Toledo Lorena A. , Shifera Amde Selassie , Santiago Esteban A.
Published: June 2024

Purpose:

The objective of this study was to present a rare case of prolonged and severe ocular monkeypox virus infection in the absence of systemic manifestations.

Methods:

This was a single case report.

Results:

A 60-year-old man, having been symptomatic for 9 days, presented with several umbilicated, ulcerated papules on the left cheek, left side of the nose, and left upper eyelid, along with marked follicular conjunctivitis and multiple conjunctival ulcerations. Two weeks after presentation, he developed an irregular, 360° circumferential opacity in the peripheral cornea that progressed to a large epithelial defect with corneal thinning. Although the initial eyelid lesions and conjunctivitis quickly resolved, the patient experienced nonresolving corneal inflammation manifest with peripheral corneal thinning, epithelial defects, and stromal keratitis. Four months after presentation, with the presumptive diagnosis of peripheral ulcerative keratitis, the patient was treated with intravenous steroids and immunosuppressive treatment, after which the ocular surface inflammation improved. However, the inflammation recurred 12 weeks later, and the patient developed severe perilimbal necrotizing conjunctivitis, followed by recurrence of ulcerated nodular eyelid lesions. Eight months after presentation, nucleic acid amplification tests from eyelid lesion swabs returned positive for nonvariola Orthopoxviruses, which led to the diagnosis of mpox. Within 2 weeks of beginning antiviral treatment with systemic tecovirimat and cidofovir and topical trifluridine, the eyelid lesions, conjunctivitis, and corneal inflammation resolved.

Conclusions:

We present an unusual and challenging case of ocular mpox with severe ocular surface inflammation including peripheral corneal thinning and epithelial defects, without systemic disease. Initiation of antiviral treatment resulted in a quick resolution of the ocular disease.
Evaluation of two-stage hepatectomy using portal vein embolization for colorectal liver metastasis: a retrospective nationwide cohort survey in Japan
Authors: Shimizu Takayuki , Aoki Taku , Ishizuka Mitsuru , Sakamoto Katsunori , Beppu Toru , Honda Goro , Kotake Kenjiro , Yamamoto Masakazu , Takahashi Keiichi , Endo Itaru , Hasegawa Kiyoshi , Itabashi Michio , Hashiguchi Yojiro , Kotera Yoshihito , Kobayashi Shin , Yamaguchi Tatsuro , Natsume Soichiro , Tabuchi Ken , Kobayashi Hirotoshi , Yamaguchi Kensei , Tani Kimitaka , Morita Satoshi , Miyazaki Masaru , Sugihara Kenichi , Ajioka Yoichi
Published: June 2024

Introduction:

Two-stage hepatectomy (TSH) enables patients to undergo surgery for colorectal liver metastasis (CRLM), which one-stage hepatectomy cannot remove. Although the outcome of TSH has been reported, there is no original report from Japan. The aim of this retrospective study was to evaluate the outcome of TSH in Japanese patients with CRLM.

Methods:

The authors conducted a retrospective cohort study using the nationwide database that included clinical information of 12 519 patients treated with CRLM between 2005 and 2017 in Japan. The primary outcome measure was overall survival. The second outcome measure was progression-free survival. Fisher’s exact test, χ2 test and Mann–Whitney U test were conducted to examine an intergroup difference. Univariate and multivariate analyses were performed using Cox regression model. Survival analysis was performed by Kaplan–Meier method and log-rank test.

Results:

Of the database, 53 patients undergoing TSH using portal vein embolization (PVE) were identified and analyzed. Their morbidity and in-hospital mortality rates at the second hepatectomy were 26.4% and 0.0%. The mean observation period was 21.8 months. The estimated 1-, 3- and 5-year overall survival rate were 92.5%, 70.8% and 34.7%. Multivariate analyses showed that more than 10 liver nodules significantly increased the mortality risk by 4.2-fold (95% CI 1.224–14.99, P= 0.023). Survival analysis revealed that repeat hepatectomy for disease progression after TSH was superior to chemotherapy in overall survival (mean: 49.6 vs. 18.7, months, P= 0.004).

Conclusion:

In the Japanese cohort, TSH was confirmed to be a safety procedure with an acceptable survival outcome. More than 10 liver nodules may be a predictor for unfavorable outcomes of patients with CRLM undergoing TSH. Furthermore, repeat hepatectomy can be a salvage treatment for resectable intrahepatic recurrence after TSH.
Unveiling advanced strategies for therapeutic stem cell interventions in severe burn injuries: a comprehensive review
Authors: Das Pratik , Pal Debajyoti , Roy Sudipta , Chaudhuri Shubhamitra , Kesh Shyam S. , Basak Piyali , Nandi Samit K.
Published: June 2024

This comprehensive review explores the complex terrain of stem cell therapies as a potential therapeutic frontier in the healing of complicated burn wounds. Serious tissue damage, impaired healing processes, and possible long-term consequences make burn wounds a complex problem. An in-depth review is required since, despite medical progress, existing methods for treating severe burn wounds have significant limitations. Burn wounds are difficult to heal because they cause extensive tissue damage. The challenges of burn injury-induced tissue regeneration and functional recovery are also the subject of this review. Although there is a lot of promise in current stem cell treatments, there are also some limitations with scalability, finding the best way to transport the cells, and finding consistent results across different types of patients. To shed light on how to improve stem cell interventions to heal severe burn wounds, this review covers various stem cell applications in burn wounds and examines these obstacles. To overcome these obstacles, one solution is to enhance methods of stem cell distribution, modify therapies according to the severity of the burn, and conduct more studies on how stem cell therapy affects individual patients. Novel solutions may also be possible through the combination of cutting-edge technologies like nanotechnology and biotechnology. This review seeks to increase stem cell interventions by analyzing present challenges and suggesting strategic improvements. The goal is to provide a more effective and tailored way to repair serious burn wounds.
Clinical characteristics, treatment and efficacy of calcaneal osteomyelitis: a systematic review with synthesis analysis of 1118 reported cases
Authors: Jiang Nan , Chen Peng , Liu Guan-Qiao , Huang Mou-Zhang , Deng Ming-Ye , Song Ming-Rui , Zhu Run-Jiu , Zhong Hong-Fa , Xiang Da-Yong , Yu Bin
Published: June 2024

Background:

Calcaneal osteomyelitis (CO) still poses great challenges to orthopedic surgeons due to the unique anatomic and functional features of the calcaneus. This study summarized the current data regarding clinical characteristics, treatment and efficacy of CO, based on an analysis of literature-reported cases.

Materials and methods:

The authors searched the PubMed, Embase, and Cochrane Library databases to find English and Chinese studies reporting on CO patients published between 2000 and 2021, with available data for synthesis analysis. The quality of the included studies was evaluated by the National Institutes of Health (NIH) assessment scale. Effective data were extracted and pooled for analysis.

Results:

Altogether 198 studies involving 1118 patients were included, with a male-to-female ratio of 2.3 (724 males and 310 females). The median age at CO diagnosis was 46 years, with a median symptom duration of 3 months. Injury-related infections (524 cases) and diabetic foot infections (336 cases) were the two most common causes, with ulcers (468 cases) and wound sinus or exudation (209 cases) being the predominant symptoms. The overall positive culture rate was 80.2%, with polymicrobial infections accounting for 18.1%. Staphylococcus aureus was the most frequently detected pathogen (42.7%), with fungal-related infections isolated in 17 cases. Although most patients received surgical interventions (96.9%), the recurrence rate was 20.1%. The incidence of infection relapse following partial calcanectomy, total calcanectomy, debridement with implantation of local antibiotics, and debridement with or without flap or skin coverage were 31.7%, 45.0%, 16.8%, and 15.1%, respectively. The overall incidence of limb amputation was 12.4%, with all-cause and CO-related mortalities of 2.8% and 0.2%, separately.

Conclusions:

CO shared similar characteristics with extremity chronic osteomyelitis, primarily affecting young males, with trauma and diabetic foot as the leading causes and Staphylococcus aureus as the most frequently detected pathogen. Despite surgery being the primary treatment modality, clinical outcomes remained unsatisfactory, marked by high rates of infection recurrence and limb amputation.
Functional Outcomes After Transanal Total Mesorectal Excision (taTME) for Rectal Cancer
Authors: Donovan Katherine F. , Lee Katherine C. , Ricardo Alison , Berger Natalie , Bonaccorso Antoinette , Alavi Karim , Zaghiyan Karen , Pigazzi Alessio , Sands Dana , DeBeche-Adams Teresa , Chadi Sami A. , McLemore Elisabeth C. , Marks John H. , Maykel Justin A. , Shawki Sherief F. , Steele Scott R. , Albert Matthew , Whiteford Mark H. , Cheng Fu-Yuan , Wexner Steven D. , Sylla Patricia
Published: June 2024

Objective:

To investigate fecal incontinence and defecatory, urinary, and sexual functional outcomes after transanal total mesorectal excision (taTME).

Background:

Proctectomy for rectal cancer may result in alterations in defecatory, urinary, and sexual function that persist beyond 12 months. The recent multicenter phase II taTME trial demonstrated the safety of taTME in patients with stage I to III tumors.

Methods:

Prospectively registered self-reported questionnaires were collected from 100 taTME patients. Fecal continence [Fecal Incontinence Quality of Life (FIQL), Wexner], defecatory function [Colorectal Functional Outcome (COREFO)], urinary function (International Prostate Symptom Score), and sexual function (Female Sexual Function Index-female, International Index of Erectile Function-male) were assessed preoperatively (PQ), 3 to 4 months postileostomy closure (FQ1), and 12 to 18 months post-taTME [postoperative questionnaire 2 (FQ2)].

Results:

Among 83 patients who responded at all 3 time points, FIQL, Wexner, and COREFO significantly worsened postileostomy closure. Between FQ1 and FQ2, FIQL lifestyle and coping, Wexner, and COREFO incontinence, social impact, frequency, and need for medication significantly improved, while FIQL depression and embarrassment did not change. International Prostate Symptom Score did not change relative to preoperative scores. For females, Female Sexual Function Index declined for desire, orgasm, and satisfaction between PQ and FQ1, and did not improve between FQ1 and FQ2. In males, International Index of Erectile Function declined with no change between FQ1 and FQ2.

Conclusions:

Although taTME resulted in initial decline in defecatory function and fecal continence, most functional domains improved by 12 months after ileostomy closure, without returning to preoperative status. Urinary function was preserved while sexual function declined without improvement by 18 months post-taTME. Our results address patient expectations and inform shared decision-making regarding taTME.
mPD5, a peripherally restricted PICK1 inhibitor for treating chronic pain
Authors: Jensen Kathrine Louise , Chistensen Nikolaj Riis , Goddard Carolyn Marie , Jager Sara Elgaard , Kanneworff Ida Buur , Jakobsen Alexander , Noes-Holt Gith , Jiménez-Fernández Lucía , Peck Emily G. , Sivertsen Line , Baro Raquel Comaposada , Houser Grace Anne , Mayer Felix Paul , Diaz-delCastillo Marta , Topp Marie Løth , Hopkins Chelsea , Thomsen Cecilie Dubgaard , Soltan Ahmed Barakat Ibrahim , Tidemand Frederik Grønbæk , Arleth Lise , Heegaard Anne-Marie , Sørensen Andreas Toft , Madsen Kenneth Lindegaard
Published: June 2024

Chronic pain is a complex, debilitating, and escalating health problem worldwide, impacting one in five adults. Current treatment is compromised by dose-limiting side effects including high abuse liability, loss of ability to function socially and professionally, fatigue, drowsiness, and apathy. PICK1 has emerged as a promising target for the treatment of chronic pain conditions. Here, we develop and characterize a cell-permeable fatty acid conjugated bivalent peptide inhibitor of PICK1 and assess its effects on acute and chronic pain. The myristoylated myr-NPEG4-(HWLKV)2, (mPD5), self-assembles into core-shell micelles that provide favourable pharmacodynamic properties and relieves ongoing and evoked mechanical hypersensitivity, thermal hypersensitivity as well as anxio-depressive symptoms in mouse models of neuropathic and inflammatory pain following subcutaneous administration. No overt no side effects were associated with mPD5 administration, and it has no effect on acute nociception. Finally, neuropathic pain is relieved far into the chronic phase (18 weeks post SNI surgery) and while the effect of a single injection ceases after a few hours, repeated administration provides pain relief lasting up to 20 hours after the last injection.
The Hippo/YAP Pathway Mediates the De-differentiation of Corneal Epithelial Cells into Functional Limbal Epithelial Stem CellsIn Vivo
Authors: Li Yijian , Ge Lingling , Ren Bangqi , Zhang Xue , Yin Zhiyuan , Liu Hongling , Yang Yuli , Liu Yong , Xu Haiwei
Published: June 2024

Regeneration after tissues injury is often associated with cell fate plasticity, which restores damaged or lost cells. The de-differentiation of corneal epithelial cells (CECs) into functional stem cells after the ablation of innate stem cells, known as limbal epithelial stem cells (LESCs), remains controversial. In this study, we showed the functional maintenance of corneal epithelium after the ablation of innate stem cells, and the regeneration of functional LESCs, which maintained corneal transparency, prevented corneal conjunctivalization and participated in the wound healing. Subsequent intravital lineage tracing revealed that CECs could de-differentiate into active or quiescent LESCs, which functioned as well as their innate counterparts. Furthermore, the de-differentiation of CECs required an intact limbal niche, and the outcome of the competition between conjunctival and corneal epithelium for the limbal niche determined whether the de-differentiation would occur or not. Mechanically, the suppression of YAP signal promoted the de-differentiation of CECs after the ablation of innate stem cells, while the persistent activation of YAP prevented the de-differentiation of CECs after an additional alkali burn to the limbal stroma. These results will pave the way for an alternative approach to treat limbal stem cell deficiency (LSCD) by modulating the de-differentiation of CECsin vivo.
More is better: A simple antibody-based strategy for recovering all major mouse brain cell types from multiplexed single-cell RNAseq samples
Authors: Pratesi Federico , Hamilton Laura K. , Avila Lopez Jessica , Aumont Anne , Avino Mariano , Singh Nishita , Arefiev Ihor , Brunet Marie A. , Fernandes Karl J. L.
Published: June 2024

Single-cell RNA sequencing (scRNAseq) is a powerful yet costly technique for studying cellular diversity within the complexity of organs and tissues. Here, we sought to establish an effective multiplexing strategy for the adult mouse brain that could allow multiple experimental groups to be pooled into a single sample for sequencing, reducing costs, increasing data yield, and eliminating batch effects. We first describe an optimized cold temperature single-cell dissociation protocol that permits isolation of a high yield and viability of brain cells from the adult mouse. Cells isolated using this protocol were then screened by flow cytometry using a panel of antibodies, allowing identification of a single antibody, anti-Thy1.2, that can tag the vast majority of isolated mouse brain cells. We then used this primary antibody against a “universal” neural target, together with secondary antibodies carrying sample-specific oligonucleotides and the BD Rhapsody single-cell system and show that multiple adult mouse brain samples can be pooled into a single multiplexed run for scRNAseq. Bioinformatic analyses enable efficient demultiplexing of the sequenced pooled brain sample, with high tagging efficiency and precise annotation and clustering of brain cell populations. The efficiency and flexibility of the cell dissociation protocol and the two-step multiplexing strategy simplifies experimental design, optimizes reagent usage, eliminates sequencing batch effects and reduces overall experimental costs.
Using a Novel Digital Go/No-Go to Dissociate Intra-subject Temporal Fluctuations in Reaction Time and Accuracy
Authors: Nguyen Theresa M. , Ross Mindy K. , Ning Emma , Kabir Sarah , Cladek Andrea T. , Barve Amruta , Kennelly Ellyn , Hussain Faraz , Duffecy Jennifer , Langenecker Scott L. , Zulueta John , Demos Alexander P. , Ajilore Olusola A. , Leow Alex D.
Published: June 2024

Impulsivity can be a risk factor for serious complications for those with mood disorders. To understand intra-individual impulsivity variability, we analyzed longitudinal data of a novel gamified digital Go/No-Go (GNG) task in a clinical sample (n=43 mood disorder participants, n=17 healthy controls) and an open-science sample (n=121, self-reported diagnoses). With repeated measurements within-subject, we disentangled two aspects of GNG: reaction time and accuracy in response inhibition (i.e., incorrect No-Go trials) with respect to diurnal and potential learning effects. Mixed-effects models showed diurnal effects in reaction time but not accuracy, with a significant effect of hour on reaction time in the clinical sample and the open-science sample. Moreover, subjects improved on their response inhibition but not reaction time. Additionally, significant interactions emerged between depression symptom severity and time-of-day in both samples, supporting that repeated administration of our GNG task can yield mood-dependent circadian rhythm-aware biomarkers of neurocognitive function.
Association between lactate to albumin ratio and acute kidney injury in patients with sepsis: a retrospective cohort study from MIMIC-IV database
Authors: Wang Yaotang , Yu Haixia
Published: June 2024

Background

The mortality rate of sepsis-associated acute kidney injury (S-AKI) is high, yet there is a lack of authoritative prognostic criteria for its outcome. The lactate-to-albumin ratio (LAR) has been associated with mortality in conditions such as sepsis, heart failure, and respiratory failure. However, there is limited research on the relationship between LAR and S-AKI. This study aimed to investigate the association between LAR and the mortality rate of S-AKI.

Methods

In our study, we performed a retrospective cohort analysis using the Medical Information Mart for Intensive Care (MIMIC)-IV database. Our primary focus was on determining the occurrence of 30-day, 60-day, and in-hospital mortality as the primary outcomes. To validate the association between LAR and these outcomes, we employed multivariable logistic Cox regression models to calculate hazard ratios.

Results

This study included a total of 4793 participants, with a mean age of 63.3 years and a median LAR (log2) of -0.90. After accounting for confounding variables, it was found that patients in the highest LAR (log2) quartile had a higher risk of mortality compared to those in the lowest LAR (log2) quartile. The adjusted hazard ratios for mortality were 1.44 (95% confidence interval [CI]: 1.36∼1.53), 1.38 (95% CI: 1.32∼1.46), and 1.47 (95% CI: 1.39∼1.56) for the highest LAR (log2) quartile, respectively. Moreover, the adjusted hazard ratios for mortality at 30-day, 90-day, and in-hospital intervals were 1.28 (95% CI: 1.23∼1.33), 1.26 (95% CI: 1.21∼1.31), and 1.27 (95% CI: 1.22∼1.32) for each 1 unit increase in LAR, respectively.

Conclusion

This study indicates that in septic AKI patients, a higher LAR is associated with an increased risk of all-cause mortality within 30-day, 90-day and in-hospital mortality. This suggests that LAR may serve as an independent risk factor for adverse outcomes in septic AKI patients.
Genetic encoding and expression of RNA origami cytoskeletons in synthetic cells
Authors: Tran Mai P. , Chakraborty Taniya , Poppleton Erik , Monari Luca , Giessler Franziska , Göpfrich Kerstin
Published: June 2024

The central dogma at the core of molecular biology states that information flows from DNA to RNA and then to protein. Our research seeks to introduce a conceptually novel approach towards synthetic life by leveraging RNA origami, as an alternative to proteins, requiring only a single copying step between genetic information and function. Here, we report the genetic encoding and expression of an RNA origami cytoskeleton-mimic within giant unilamellar lipid vesicles (GUVs). We design the first RNA origami tiles which fold co-transcriptionally from a DNA template and self-assemble into higher-order 3D RNA origami nanotubes at constant 37◦C in GUVs, where they reach several micrometers in length. Unlike pre-formed and encapsulated DNA cytoskeletons, these GUVs produce their own molecular hardware in an out-of-equilibrium process fuelled by nucleotide feeding. To establish genotype-phenotype correlations, we investigate how sequence mutations govern the contour and persistence length of the RNA origami nanotubes with experiments and coarse-grained molecular-dynamics simulations, realizing a phenotypic transition to closed rings. Finally, we achieve RNA origami cortex formation and GUV deformation without chemical functionalization by introducing RNA aptamers into the tile design.Altogether, this work pioneers the expression of RNA origami-based hardware in vesicles as a new approach towards active, evolvable and RNA-based synthetic cells.
LDL exposure disrupts mitochondrial function and dynamics in a hippocampal neuronal cell line
Authors: Farias Hémelin Resende , Ramos Jessica Marques Obelar , Griesang Caroline , Santos Lucas , Ramires Junior Osmar Vieira , de Souza Debora Guerini , Ferreira Fernanda Silva , Somacal Sabrina , Martins Leo Anderson Meira , de Souza Diogo Onofre Gomes , Moreira José Cláudio Fonseca , de Souza Wyse Ângela Terezinha , Guma Fátima Theresinha Costa Rodrigues , de Oliveira Jade
Published: June 2024

Hypercholesterolemia has been associated with cognitive dysfunction and neurodegenerative disease. Moreover, this metabolic condition disrupts the blood-brain barrier, allowing Low-Density Lipoprotein (LDL) to enter the Central Nervous System. Thus, we investigated the effects of LDL exposure on mitochondrial function in a mouse hippocampal neuronal cell line (HT-22). HT-22 cells were exposed to human LDL (50 and 300 μg/mL) for 24 hours. After this, intracellular lipid droplet (LD) content, cell viability, cell death, and mitochondrial parameters were performed. We found that the higher LDL concentration LDL increases LD content compared to control. Both concentrations increased the number of Annexin V-positive cells, indicating apoptosis. Moreover, in mitochondrial parameters, the exposure of LDL on hippocampal neuronal cell line leads to a decrease in mitochondrial complexes I and II in both concentrations tested and a reduction in Mitotracker™ Red fluorescence and Mitotracker™ Red and Mitotracker™ Green ratio in the higher concentration, indicating dysfunction in the mitochondria. The LDL incubation induces mitochondrial superoxide production and a decrease in superoxide dismutase activity in the lower concentration in HT-22 cells. Finally, hippocampal neuronal cell line exposed to LDL exhibit an increase in the expression of genes associated with mitochondrial fusion (OPA1 and Mitofusin 2) in the lower concentration. In conclusion, our findings suggest that LDL exposure induces mitochondrial dysfunction and modulation in mitochondrial dynamics in the hippocampal neuronal cells.
Molecular insights into the interaction between a disordered protein and a folded RNA
Authors: Mitra Rishav , Usher Emery T. , Dedeoğlu Selin , Crotteau Matthew J. , Fraser Olivia A. , Yennawar Neela H. , Gadkari Varun V. , Ruotolo Brandon T. , Holehouse Alex S. , Salmon Loïc , Showalter Scott A. , Bardwell James C. A.
Published: June 2024

Intrinsically disordered protein regions (IDRs) are well-established as contributors to intermolecular interactions and the formation of biomolecular condensates. In particular, RNA-binding proteins (RBPs) often harbor IDRs in addition to folded RNA-binding domains that contribute to RBP function. To understand the dynamic interactions of an IDR-RNA complex, we characterized the RNA-binding features of a small (68 residues), positively charged IDR-containing protein, SERF. At high concentrations, SERF and RNA undergo charge-driven associative phase separation to form a protein- and RNA-rich dense phase. A key advantage of this model system is that this threshold for demixing is sufficiently high that we could use solution-state biophysical methods to interrogate the stoichiometric complexes of SERF with RNA in the one-phase regime. Herein, we describe our comprehensive characterization of SERF alone and in complex with a small fragment of the HIV-1 TAR RNA (TAR) with complementary biophysical methods and molecular simulations. We find that this binding event is not accompanied by the acquisition of structure by either molecule; however, we see evidence for a modest global compaction of the SERF ensemble when bound to RNA. This behavior likely reflects attenuated charge repulsion within SERF via binding to the polyanionic RNA and provides a rationale for the higher-order assembly of SERF in the context of RNA. We envision that the SERF-RNA system will lower the barrier to accessing the details that support IDR-RNA interactions and likewise deepen our understanding of the role of IDR-RNA contacts in complex formation and liquid-liquid phase separation.

SIGNIFICANCE

Subcellular organization through the formation of biomolecular condensates has emerged as an important contributor to myriad cellular functions, with implications in homeostasis, stress response, and disease. To understand the general and specific principles that support condensate formation, we must interrogate the interactions and assembly of their constituent biomolecules. To this end, this study introduces a simple model system comprised of a small, disordered protein and small RNA that undergo charge-driven, associative phase separation. In addition to extensive biophysical characterization of these molecules and their complex, we also generate new insights into mode of interaction and assembly between an unstructured protein and a structured RNA.
Isoform and pathway-specific regulation of post-transcriptional RNA processing in human cells
Authors: Bedi Karan , Magnuson Brian , Narayanan Ishwarya Venkata , McShane Ariel , Ashaka Mario , Paulsen Michelle T. , Wilson Thomas E. , Ljungman Mats
Published: June 2024

Steady-state levels of RNA transcripts are controlled by their rates of synthesis and degradation. Here we used nascent RNA Bru-seq and BruChase-seq to profile RNA dynamics across 16 human cell lines as part of ENCODE4 Deeply Profiled Cell Lines collection. We show that RNA turnover dynamics differ widely between transcripts of different genes and between different classes of RNA. Gene set enrichment analysis (GSEA) revealed that transcripts encoding proteins belonging to the same pathway often show similar turnover dynamics. Furthermore, transcript isoforms show distinct dynamics suggesting that RNA turnover is important in regulating mRNA isoform choice. Finally, splicing across newly made transcripts appears to be cooperative with either all or none type splicing. These data sets generated as part of ENCODE4 illustrate the intricate and coordinated regulation of RNA dynamics in controlling gene expression to allow for the precise coordination of cellular functions.
SEGS-1 episomes generated during cassava mosaic disease influence disease severity
Authors: Chiunga Evangelista , Aimone Catherine D. , Rajabu Cyprian , Dallas Mary M. , Ndunguru Josep , Ascencio-Ibáñez José T. , Ateka Elijah M. , Hanley-Bowdoin Linda
Published: June 2024

Cassava is an important root crop that is produced by smallholder farmers across Sub-Saharan Africa. Cassava mosaic disease (CMD), which is caused by a complex of cassava mosaic begomoviruses (CMBs), is one of the most devastating diseases of cassava. A previous study showed that SEGS-1 (sequences enhancing geminivirus symptoms), which occurs in both the cassava genome and as an episome during CMD, can increase CMD disease severity and overcome host resistance. In this report, we examined the effects of exogenously applied SEGS-1 on the incidence of CMB infection, symptom severity, and viral DNA copy number in five cassava cultivars that ranged from highly susceptible to highly resistant to CMD. These studies revealed that the effect of SEGS-1 is cultivar dependent. Susceptible cultivars developed severe CMD in the absence or presence of exogenous SEGS-1, while exogenous SEGS-1 increased disease severity in cultivars carrying CMD2 but not CMD1 resistance. Analysis of infected plants in the absence of exogenous SEGS-1 revealed that some, but not all cultivars, form SEGS-1 episomes during CMD. The presence of endogenous SEGS-1 episomes in TME14, a CMD2 resistant cultivar, correlated with CMD severity. In contrast, TME3, a closely related CMD2 cultivar, did not produce endogenous SEGS-1 episomes and showed more resistance than TME14. DNA sequence analysis indicated that the different capacities of TME3 and TME14 to form SEGS-1 episomes is unlikely due to sequence differences in and around their genomic SEGS-1 loci. Because of its inability to form episomes, TME3 was used to map the functional regions of SEGS- 1 to sequences flanking the epsiome junction, but junction itself was not required for activity. Together, these experiments provided insight into the functional form of SEGS-1 in cassava and the effect of cassava genotype on SEGS-1 activity.
Improving Adherence to electronic Patient-Reported Outcomes:Study Protocol for a Systematic Literature Review and subsequent qualitative Interviews
Authors: Mukowski-Kickhöfel Rebecca , Otto Lisa , Rogge Alizé
Published: June 2024

Background

Electronic patient-reported outcomes (ePROs) are frequently used to include the patients’ perspective in clinical trials and routine clinical care. In contrast to data collected in clinical trials, PRO response rates tend to decrease drastically overtime in routine clinical care, leading to missing data and questionable validity of results reported. This study aims to investigate factors influencing patient adherence to digitally collected PROs.

Method

The study comprises of three steps: (1) a systematic literature review (SLR) focusing on factors increasing PRO response rates, (2) focus group interviews with patients to develop criteria on how to increase response rates for real-world evidence (RWE), and subsequently, (3) an anonymous online survey to evaluate developed criteria. The SLR will follow the PRISMA guidelines. The inclusion criteria for the review encompass studies focusing exclusively on patients or people recruited via medical personnel. The literature review will include studies from various settings, encompassing publications in German or English language published within the past 10 years (2014-2024). Participants for the focus group interviews (n = 6-8/group) will be recruited via patient advisory boards or groups/organisations. The findings of the SLR and the focus group form the basis for the anonymous online survey to include more patients.

Outlook

The results of the study can be incorporated into the development and implementation of digital collection methods for PROs, with the aim of improving adherence to PROs and thus robustness of RWE data.
Identification of perturbation-responsive regions and genes in comparative spatial transcriptomics atlases
Authors: Teo Alan Yue Yang , Gautier Matthieu , Brock Laurent , Tsai Jennifer Y. J. , de Coucy Alexandra , Laskaratos Achilleas , Regazzi Nicola , Barraud Quentin , Sofroniew Michael V. , Anderson Mark A. , Courtine Grégoire , Squair Jordan W. , Skinnider Michael A.
Published: June 2024

We introduce Vespucci, a machine-learning method to identify perturbation-responsive regions, genes and gene programs within comparative spatial transcriptomics atlases. We validate Vespucci on simulated and published datasets and show that it outperforms 19 published computational methods for spatial transcriptomics. We apply Vespucci to expose the spatial organization of gene programs activated by therapies that guide repair of the injured spinal cord.