Conformation-Dependent Hydrogen-Bonding Interactions in a Switchable Artificial Metalloprotein.

Hydrogen-bonding (H-bonding) interactions in metalloprotein active sites can critically regulate enzyme function. Changes in the protein structure triggered by interplay with substrates, products, and partner proteins are often translated to the metallocofactor by way of specific changes in H-bond networks connected to the active site. However, the complexities of metalloprotein architecture and mechanism often preclude our ability to define the precise molecular interactions giving rise to these intricate regulatory pathways. To address this shortcoming, we have developed conformationally switchable artificial metalloproteins (swArMs) in which allosteric Gln-binding triggers protein conformational changes that impact the microenvironment surrounding an installed metallocofactor. Herein, we report a combined structural, spectroscopic, and computational approach to enhance the conformation-dependent changes in H-bond interactions surrounding the metallocofactor site of a swArM. Structure-informed molecular dynamics simulations were employed to predict point mutations that could enhance active site H-bond interactions preferentially in the Gln-bound holo-conformation of the swArM. Testing our predictions via the unique infrared spectral signals associated with the metallocofactor site, we have identified three key residues capable of imparting conformational control over the metallocofactor microenvironment. The resultant swArMs not only model biologically relevant structural regulation but also provide an enhanced Gln-responsive biological probe to be leveraged in future biosensing applications.

Probing calmodulin-NO synthase interactions via site-specific infrared spectroscopy: an introductory investigation.

Calmodulin (CaM) binds to a linker between the oxygenase and reductase domains of nitric oxide synthase (NOS) to regulate the functional conformational dynamics. Specific residues on the interdomain interface guide the domain-domain docking to facilitate the electron transfer in NOS. Notably, the docking interface between CaM and the heme-containing oxygenase domain of NOS is isoform specific, which is only beginning to be investigated. Toward advancing understanding of the distinct CaM-NOS docking interactions by infrared spectroscopy, we introduced a cyano-group as frequency-resolved vibrational probe into CaM individually and when associated with full-length and a bi-domain oxygenase/FMN construct of the inducible NOS isoform (iNOS). Site-specific, selective labeling with p-cyano-L-phenylalanine (CNF) by amber suppression of CaM bound to the iNOS has been accomplished by protein coexpression due to the instability of recombinant iNOS protein alone. We introduced CNF at residue 108, which is at the putative CaM-heme (NOS) docking interface. CNF was also introduced at residue 29, which is distant from the docking interface. FT IR data show that the 108 site is sensitive to CaM-NOS complex formation, while insensitivity to its association with the iNOS protein or peptide was observed for the 29 site. Moreover, narrowing of the IR bands at residue 108 suggests the C≡N probe experiences a more limited distribution of environments, indicating side chain restriction apparent for the complex with iNOS. This initial work sets the stage for residue-specific characterizations of structural dynamics of the docked states of NOS proteins.

Socio-cultural determinants of tobacco consumption: a study of migrant and local workers in a district of Punjab, India.

Although the use of tobacco in the state of Punjab is below the national statistics of India, according to Global Adult Tobacco Survey (GATS) 2016 Report, its consumption has increased. Also, what sets it apart from rest of the nation is the cultural as well as the religious taboo associated with tobacco use. The case of Punjab is also peculiar because of the socio demographic profile of the region which has changed since Green Revolution. Having said that, use of tobacco is a social phenomenon and it becomes important to place tobacco use in a social and cultural context. The aim of the study is to explore the socio -cultural determinants of tobacco use within local and migrant groups in a district of Punjab. The tobacco users in the district were divided into groups of local and migrant labourers and focus group discussion was carried out with them. There were various social correlates of tobacco use in the district which saw variations within and across these groups. The socio demographic factors of age, gender, religion and place of residence were related to the use of tobacco and presented similarities and variations across the groups. The study concluded that tobacco prevention and control policies should include cultural interventions to change current traditional practices and social determinants which influence the use of tobacco.

Solvent effect on the competition between weak and strong interactions in phenol solutions studied by near-infrared spectroscopy and DFT calculations.

Near-infrared (NIR) spectra of phenol in a series of non-aromatic and aromatic solvents were recorded to study the competition between various types of solute-solute and solute-solvent interactions. Depending on the phenol concentration, the free OH and OH involved in the OH⋯OH interactions in the dimers and higher associates are present in cyclohexane solutions. On the other hand, free OH does not appear in Cl-containing solvents since at a low phenol content the OH groups participate in the OH⋯Cl interactions. In CCl4 and tetrachloroethylene this interaction is weak, while in chlorobenzene the strength of this interaction is higher. In the aromatic solvents the solute-solute OH⋯OH interactions compete with the solute-solvent OH⋯π and aromatic CH⋯OH ones. Consequently, the degree of self-association of phenol in aromatic solvents is smaller than that in non-aromatic ones. The strength of the OH⋯π interactions increases with growing electron-donating ability of the substituents in the benzene derivatives. This observation obtained from the NIR spectra is in line with the results of the theoretical calculations (DFT). A clear correlation appears between the number of methyl groups in aromatic solvents and the population of the free OH groups. The methyl groups are steric hindrances and impede the formation of the OH⋯OH and OH⋯π interactions. Our results suggest the presence of aromatic CH⋯OH solute-solvent interactions, not observed in previous studies. NIR spectroscopy appears to be a powerful tool for exploration of free and weakly-bonded OH groups.

Post-operative Patellar Tilt More than 10° Can Affect Certain Components of Knee Society Score After Total Knee Arthroplasty at 2-Year Follow-Up.

Introduction: The effect of post-operative patella tilt on functional outcomes after total knee arthroplasty remains unclear. Our study aimed to analyze the relationship of post-operative patellar tilt with functional outcome scores after total knee arthroplasty. Materials and Methods: Patient data were retrieved from our institution's prospectively maintained total knee arthroplasty. Three hundred three patients who underwent unilateral TKA from Jan 2012 to March 2017 were included in the study. After excluding patients with incomplete and lost follow-up data, 213 patients were analyzed. Radiographs of pre-operative and post-operative skyline views were used for patella tilt and patella displacement measurement at pre-op, post-op 1 year, and post-op 2 years. Three functional outcome scoring systems, SF-36, KSS, and WOMAC, were applied for function evaluation at different post-operative time points. Patients were divided into three subgroups according to the patella tilt, which includes less than 5°, 5.1-10°, and more than 10°. Statistical analysis was done to identify the relationship between patella tilt and functional outcomes. Results: Mean post-operative patella tilt was significantly lower than the mean pre-operative patella tilt (3.35 ± 3.91 vs. 5.65 ± 4.41, p < 0.001). There was no significant difference in patella displacement among pre- and post-operative status. KSS functional score was significantly poor at post-op 1 year and KSS objective score at post-op 2 years in patients with more than 10° patella tilt. SF-36 and WOMAC were not significantly different among the groups. There was no significant difference in post-operative function between non-resurfaced and resurfaced patella patients evaluated with three scoring systems. Conclusion: We have found significantly less post-operative patella tilt after TKA than pre-operative patella tilt with or without patella resurfacing. Increased post-operative patella tilt of more than 10° can affect specific functional outcomes. Patella resurfacing does not affect the post-operative functional outcome compared to non-resurfacing of the patella post-op 2 years. Level of Evidence: III. Supplementary Information: The online version contains supplementary material available at 10.1007/s43465-023-01077-0.

Oral health assessment and treatment needs of 12- and 15-year-old children residing in tribal welfare and private hostels - A comparative study.

Background and Objectives: Oral health is an integral part of general health. Providing oral health care facilities and creating awareness about the oral health problem in under-served children and communities may lead to a better oral-health-related quality of life of the individual. The present study aims to assess and compare the oral health status and treatment needs of 12- and 15-year-old children residing in tribal welfare hostels and other private hostels of Bhopal district, Madhya Pradesh. Materials and Methods: The present descriptive cross-sectional study consisted of a total sample size of 800 children, 400 in each group (tribal welfare hostel and private hostel groups). Oral health status and treatment needs were assessed using World Health Organization proforma 1997. Oral health behavior including health of teeth and gums, oral hygiene aids, brushing frequency, consumption of sweets in between meals, and present general and oral health was assessed. Utilization of dental services was also assessed using a pre-designed questionnaire, which was completed by the study participants. Results: Statistical analysis was carried out using Chi-square test. Significant differences were noted between the groups in regarding oral health behaviors and visit to a dentist during the past 12 months (P = 0.002*), which were lower in tribal children. Tribal children were having higher dental fluorosis as compared to the private hostel children (P = 0.043*). Decay in permanent teeth (P = 0.006*) and missing of permanent teeth (P = 0.05*) were higher among tribal children. Conclusion: The present study revealed a poor oral health status and treatment needs of tribal children. Tribal children were having higher dental fluorosis as compared to the private hostel children. Decay in permanent teeth and missing of permanent teeth were higher among tribal children. Oral health behaviors and utilization of dental services were lower in tribal children. Good oral health has a definitive influence on general health and thus contributes to self-image and social interaction.

Patient-Reported Outcomes Following Open Broström-Gould Procedure: Minimum 5-Year Follow-Up.

Background: Ankle sprains are a common musculoskeletal injury among the general population and often involve the lateral ligament complex. Although the majority of ankle sprains are treated successfully with nonsurgical conservative measures, an estimated 5% to 20% of ankle injuries ultimately develop chronic lateral ankle instability (CAI). Multiple surgical treatment modalities for the lateral ankle complex exist, such as anatomical and nonanatomical reconstruction. The current gold standard for primary surgical repair is the Broström-Gould procedure. This is the first article to provide PROMIS scores following BG and the largest study with 5-year outcomes for an open BG. Methods: This was a descriptive study of a retrospective cohort of patients undergoing a BG with a minimum follow-up of 5 years. Patient-reported outcome instruments collected postoperatively were PROMIS Pain, Physical Function, Depression, and FAAM. Further preoperative clinic characteristics were analyzed to correlate with the final outcome. The electronic medical record was queried for Current Procedural Terminology (CPT) code 27698 (Broström-Gould) from January 2010 to June 2017. Surveys were conducted in the clinic and through phone interviews. Patient charts were reviewed to obtain basic patient demographic information including sex, age, race, and body mass index (BMI). The following preoperative variables were recorded: history of prior CAI procedures, history of major trauma, duration of symptoms, number of diagnosed ankle sprains, other collagen pathologies, generalized ligament laxity, participation in sports/activity level, peroneal subluxation, clinically diagnosed peroneus longus or brevis tendinopathy, deltoid ligament injury, anterior ankle impingement, and posterior ankle impingement. The PROMIS and Foot and Ankle Ability Measure (FAAM) scores were obtained with a combination of clinic and phone interviews. Data were aggregated in Microsoft Excel and entered in R (version 4.2.0) for statistical analysis. Results: Our results show that the minimum 5-year patient-reported PROMIS scores for patients following a Broström-Gould procedure are as follows: PROMIS physical function, 50.5; PROMIS pain interference, 48.2; and PROMIS depression, 38.2. This indicates, at a minimum, that patients 5 years removed from the procedure are within 1 SD of the general population in regard to PROMIS physical function and pain. Our patient-reported FAAM, activities of daily living, and FAAM sports scores were 59.6 and 13.0 respectively. Preoperative magnetic resonance imaging (MRI) findings were recorded. Arthroscopic examination was performed before lateral ligaments reconstruction for patients with intra-articular pathologies confirmed on MRI. Conclusion: The findings from our study offer evidence supporting the effectiveness of the Broström-Gould procedure to be associated with normal physical function, even 5 years after surgery. Furthermore, our research identified specific factors such as tobacco use, diabetes, and sports participation that independently correlated with reported outcome measures. These insights enable physicians to better manage patient expectations and tailor treatment strategies accordingly. Our study establishes a foundation for future prospective research endeavors that aim to leverage the PROMIS system for comprehensive outcome assessments. Level of Evidence: Level III, retrospective cohort study.

Anatomical Structures at Risk in Percutaneous Distal Bunionette Correction.

Bunionette deformity is an incredibly pervasive issue in our society with almost a quarter of individuals being affected by it. As it is so common, there are numerous techniques and approaches to correct the deformity. Currently, there is a growing trend that favors percutaneous osteotomy of the bunionette. As there are multiple osteotomy sites, there are anatomical considerations that must be made at each one. The purpose of this study was to investigate the anatomic structures at risk during distal osteotomy of bunionette deformity using a Shannon burr. Using 11 fresh cadaver specimens, the fifth metatarsal was accessed through a carefully marked portal. A Shannon burr was employed for the osteotomy. Dissections were performed to assess potential damage to critical structures, including the lateral dorsal cutaneous nerve (LDCN), abductor digiti minimi (ADM), and extensor digitorum longus (EDL). Measurements were taken from the osteotomy site to each structure. The distal osteotomy site was on average greater than 8 mm from the EDL and ADM, whereas it was 1.64 mm from the LDCN. The Shannon burr made contact with and transected the LDCN on 2 occasions. However, previous studies have highlighted potential anatomical variations of the LDCN that arise distally. The study underscored the challenges posed by minimally invasive approaches to treating bunionette deformity and highlighted the need for cautious consideration when using percutaneous methods.Level of Clinical Evidence: 5.

Nanophytosomal Gel of Heydotis corymbosa (L.) Extract against Psoriasis: Characterisation, In Vitro and In Vivo Biological Activity.

The current study was conducted to examine the possible advantages of Heydotis corymbosa (L.) Lam. extract nanogel as a perspective for enhanced permeation and extended skin deposition in psoriasis-like dermatitis. Optimised nanophytosomes (NPs) were embedded in a pluronic gel base to obtain nanogel and tested ex vivo (skin penetration and dermatokinetics) and in vivo. The optimised NPs had a spherical form and entrapment efficiency of 73.05 ± 1.45% with a nanosized and zeta potential of 86.11 nm and -10.40 mV, respectively. Structural evaluations confirmed encapsulation of the drug in the NPs. Topical administration of prepared nanogel to a rat model of psoriasis-like dermatitis revealed its specific in vivo anti-psoriatic efficacy in terms of drug activity compared to the control and other formulations. Nanogel had improved skin integrity and downregulation of inflammatory cytokines. These findings suggest that developed phytoconstituent-based nanogel has the potential to alleviate psoriasis-like dermatitis with better skin retention and effectiveness.

Anatomical Structures at Risk in Percutaneous Preparation Talonavicular Fusion.

BACKGROUND: The stability of the hindfoot greatly relies on the integrity of the talonavicular joint. Pathologies affecting this joint often necessitate fusion. Our study explores the risks posed to neurovascular and tendon structures during simulated percutaneous talonavicular joint preparation for fusion. METHODS: In 9 fresh cadaver specimens, the talonavicular joint was accessed through two portals. A 2-mm Shannon burr was employed for joint surface preparation with distraction provided by a pin-based distractor. Dissections were performed to assess potential damage to critical structures, including the dorsalis pedis artery, superficial and deep peroneal nerves, extensor hallucis longus (EHL), and tibialis anterior (TA) tendons. RESULTS: The dorsal portal site was found to be significantly closer to important structures compared to the medial portal site. The Shannon burr made contact with various structures, with a single transection identified for both deep and superficial peroneal nerve branches. During the dorsal portal site approach, potential injury to the EHL tendon was identified as concern. CONCLUSION: This study sheds light on the potential risks associated with percutaneous dorsal and medial joint preparation approaches using a Shannon burr.Level of Evidence:Level V, mechanism-based reasoning..

SKP2 Knockout in Rb1/p53-Deficient Mouse Models of Osteosarcoma Induces Immune Infiltration and Drives a Transcriptional Program with a Favorable Prognosis.

Osteosarcoma is an aggressive bone malignancy with a poor prognosis. One putative proto-oncogene in osteosarcoma is SKP2, encoding a substrate recognition factor of the SCF E3 ubiquitin ligase. We previously demonstrated that Skp2 knockout in murine osteosarcoma improved survival and delayed tumorigenesis. Here, we performed RNA sequencing (RNA-seq) on tumors from a transgenic osteosarcoma mouse model with conditional Trp53 and Rb1 knockouts in the osteoblast lineage ("DKO": Osx1-Cre;Rb1lox/lox;p53lox/lox) and a triple-knockout model with additional Skp2 germline knockout ("TKO": Osx1-Cre;Rb1lox/lox;p53lox/lox;Skp2-/-), followed by qPCR and immunohistochemistry validation. To investigate the clinical implications of our results, we analyzed a human osteosarcoma patient cohort ("NCI-TARGET OS") with RNA-seq and clinical data. We found large differences in gene expression after SKP2 knockout. Surprisingly, we observed increased expression of genes related to immune microenvironment infiltration in TKO tumors, especially the signature genes for macrophages and to a lesser extent, T cells, B cells, and vascular cells. We also uncovered a set of relevant transcription factors that may mediate these changes. In osteosarcoma patient cohorts, high expression of genes upregulated in TKO was correlated with favorable overall survival, which was largely explained by the macrophage gene signatures. This relationship was further supported by our finding that SKP2 expression was negatively correlated with macrophage infiltration in the NCI-TARGET osteosarcoma and the TCGA Sarcoma cohorts. Overall, our findings indicate that SKP2 may mediate immune exclusion from the osteosarcoma tumor microenvironment, suggesting that SKP2 modulation in osteosarcoma may induce antitumor immune activation.

Artificial intelligence assisted preoperative planning and 3D-printing guiding frame for percutaneous screw reconstruction in periacetabular metastatic cancer patients.

Background: The percutaneous screw reconstruction technique, known as the "Tripod Technique," has demonstrated favorable clinical outcomes in the management of metastatic periacetabular lesions, as evidenced by our prior investigations and corroborated by independent studies. Nevertheless, there is a steep learning curve in handling this technique, with possible complications such as intraarticular screw placement. Methods: Preoperative pelvic CT scans were acquired before surgery and utilized for the guiding frame design. A convolutional neural network model was trained with annotated data to identify the starting point and trajectory of each potential screw. A model boundary intersection detection technology was used to determine the optimal diameter and length of each screw. A non-rigid registration technology was matched with a prefabricated model of the body surface to design personalized anchoring skin pads. Finally, a polylactic acid-based guiding frame for intraoperative was custom-made with a 3D printer. Results: 12 patients underwent a guiding frame-assisted Tripod procedure for treatment of periacetabular metastatic lesions. An intraoperative CT scan was performed in all cases to confirm screw trajectories. Among 36 screws that were implanted, 26 screws were implanted as designed. The remaining ten screws drifted, but all remained within the intra-osseous conduit without any complications. The mean surgical time was 1.22 h with the guiding frame compared with 2.3 h without the guiding frame. Following the surgical procedure, a noteworthy enhancement in pain management, as evidenced by a reduction in scores on the visual analog scale (p < 0.01), and an improvement in functional status, as assessed through the Eastern Cooperative Oncology Group score (p < 0.01), were observed when compared to the patient's pre-operative condition. Conclusion: This proof-of-concept investigation demonstrates that the amalgamation of AI-assisted surgical planning and additive manufacturing can improve surgical accuracy and shorten surgical duration. While access to this technology is currently constrained during its early stages of development, it is anticipated that these limitations will diminish as the potential of AI and additive manufacturing in facilitating complex orthopedic procedures becomes more evident, leading to a surge in interest and adoption of this approach.

Targeted inhibition of SCFSKP2 confers anti-tumor activities resulting in a survival benefit in osteosarcoma.

Osteosarcoma(OS) is a highly aggressive bone cancer for which treatment has remained essentially unchanged for decades. Although OS is characterized by extensive genomic heterogeneity and instability, RB1 and TP53 have been shown to be the most commonly inactivated tumor suppressors in OS. We previously generated a mouse model with a double knockout (DKO) of Rb1 and Trp53 within cells of the osteoblastic lineage, which largely recapitulates human OS with nearly complete penetrance. SKP2 is a repression target of pRb and serves as a substrate recruiting subunit of the SCFSKP2 complex. In addition, SKP2 plays a central role in regulating the cell cycle by ubiquitinating and promoting the degradation of p27. We previously reported the DKOAA transgenic model, which harbored a knock-in mutation in p27 that impaired its binding to SKP2. Here, we generated a novel p53-Rb1-SKP2 triple-knockout model (TKO) to examine SKP2 function and its potential as a therapeutic target in OS. First, we observed that OS tumorigenesis was significantly delayed in TKO mice and their overall survival was markedly improved. In addition, the loss of SKP2 also promoted an apoptotic microenvironment and reduced the stemness of DKO tumors. Furthermore, we found that small-molecule inhibitors of SKP2 exhibited anti-tumor activities in vivo and in OS organoids as well as synergistic effects when combined with a standard chemotherapeutic agent. Taken together, our results suggest that SKP2 inhibitors may reduce the stemness plasticity of OS and should be leveraged as next-generation adjuvants in this cancer.

Management of partially absorbed white soft cataract post penetrating injury to eye.

Background: A measurable burden to the emergency ophthalmology department is represented by ocular trauma in pediatric patients. Traumatic cataracts still result in visual disability despite great advancements in diagnostic and treatment methods. Cataract surgery with intraocular lens (IOL) implantation aids in the improvement of visual acuity in such cases. Duration of trauma is an important prognostic factor for recovery of visual acuity before amblyopia sets in young patients with penetrating ocular injury. Purpose: This video deals with the management of a case of partially absorbed traumatic cataract in a scenario of an old and neglected penetrating injury. This case had a corneal scar, ruptured anterior lens capsule, and posterior synechiae formation between the posterior pigmented epithelium of the iris and the lens capsule. Synopsis: In a case of penetrating ocular injury, one should always suspect violation of posterior lens capsule, weakened or broken zonules and retained intraocular foreign body. In this case, a circular capsulorhexis is difficult to attain. After staining the capsule with trypan blue dye, viscoelastic substance is instilled in the anterior chamber to have good control over the rhexis and to avoid rhexis run out. In case the rhexis runs off to the equator, a pair of Vannas scissors is used to cut the extended flap. The cataract is partially absorbed, white and soft in nature and is easily mobilized from the bag and eaten up via phacoaspiration. Before implantation of posterior chamber intraocular lens (PCIOL) in the sulcus, posterior synechiae are released by swiping a cyclodialysis spatula in the sulcus area. Visual axis is cleared by giving nicks in the posterior capsule to remove the central dense posterior plaque. Automated anterior vitrectomy is done and a three-piece PCIOL is implanted safely in the ciliary sulcus. Retained viscoelastic substance is washed, intracameral antibiotic is instilled, and the anterior chamber is subsequently formed via stromal wound hydration. Highlights: Through this video, we tried to show how one should proceed with phacoaspiration with intraocular lens implantation in a case of traumatic cataract post penetrating injury in a sequential manner. Video Link: https://youtu.be/20DbYUn_Fd8.

Morphological and antioxidant responses of Cicer arietinum L. genotypes exposed to combination stress of anthracene and sodium chloride.

Various mutagenic, carcinogenic pollutants such as Polycyclic Aromatic Hydrocarbons (PAHs) are released into the environment posing a negative effect on plant metabolism. All the pollutants that are emitted into the atmosphere, ultimately find their way into the plant. Soil salinity stress is one of the major determinants of crop productivity. Different plants respond differently to different abiotic stress present alone or in combination. One such combination of abiotic stress is PAHs and salinity stress. The present research aims to study the effect of the application of NaCl and Anthracene alone and in various combinations on two chickpea genotypes GPF2 and PDG4. A 21 days laboratory experiment was conducted in petriplates and growth pouches. Different concentrations of NaCl and Anthracene were given to two chickpea genotypes viz. GPF2 and PDG4, alone as well as in combinations to study morphological, physiological and antioxidant responses. Results obtained were further analyzed by using various statistical measures such as Principle Component Analysis and Two-way ANOVA. Results indicated that under the dual presence of NaCl and Anthracene, GPF2 exhibited higher activities of antioxidant enzymes and was shown to have a negative correlation with plant height and chlorophyll content. Based on the results of the present investigation, it was concluded that GPF2 was a better performing chickpea genotype towards the combined presence of Anthracene and NaCl as compared to PDG4.

Factors Associated With Poor Patient-Reported Outcomes in Isolated Gastrocnemius Recession for Heel Pain.

Background: Gastrocnemius recession is commonly performed for a variety of pathologies of the foot and ankle, yet studies characterizing risk factors associated with patient-reported outcomes are limited. In this cohort study, patient outcomes were compared against the general population for PROMIS scores with correlation analysis comparing demographics and comorbidities. Our primary goal in this study is to identify risk factors associated with poor patient-reported outcomes following isolated gastrocnemius recession for patients with plantar fasciitis or insertional Achilles tendinopathy. Methods: A total of 189 patients met inclusion criteria. The open Strayer method was preferred. However, if the myotendinous junction could not be adequately visualized without expanding the excision, then a Baumann procedure was performed. The decision between the two did not depend on preoperative contracture. Patient demographics and visual analog scale (VAS) scores were obtained via the electronic medical record. Telephone interviews were completed to collect postoperative Patient-Reported Outcomes Measurement Information System (PROMIS) and Foot Function Index (FFI) scores. The data were analyzed using the type 3 SS analysis of variance test to identify individual patient factors associated with reduced PROMIS, FFI, and VAS scores. Results: No demographic variables were found to be significantly associated with postoperative complications. Patients who reported tobacco use at the time of surgery had significantly decreased postoperative PROMIS physical function (P = .01), PROMIS pain interference (P < .05), total FFI scores (P < .0001), and each individual FFI component score. Patients undergoing their first foot and ankle surgeries reported numerous significant postoperative outcomes, including decreased PROMIS pain interference (P = .03), higher PROMIS depression (P = .04), and lower FFI pain scores (P = .04). Hypertension was significantly associated with an increased FFI disability score (P = .03) and, along with body mass index (BMI) >30 (P < .05) and peripheral neuropathy (P = .03), significantly higher FFI activity limitation scores (P = .01). Pre- and postoperative VAS scores demonstrated improvement in patient-reported pain from a mean of 5.53 to 2.11, respectively (P < .001). Conclusion: We found in this cohort that numerous patient factors were independently associated with differences in patient-reported outcomes following a Strayer gastrocnemius recession performed for plantar fasciitis or insertional Achilles tendinopathy. These factors include, but are not limited to, tobacco use, prior foot and ankle surgeries, and BMI. This study strengthens previous reports demonstrating the efficacy of isolated gastrocnemius recession and elucidates variables that may affect patient-reported outcomes. Level of Evidence: Level III, retrospective cohort study.

Monitoring structural fluctuations of discotic liquid crystal during phase transitions.

The discotic liquid crystal 4-((2, 3, 4-tris (octyloxy) phenyl) diazenyl) benzoic acid, hereafter referred as DLC A8, exhibited in dimeric form has been studied using a combination of quantum chemical approaches and vibrational spectroscopy. This study investigates the structural alteration of DLC A8 associated with phase transition. The phase transitions of DLC A8 are Iso â†’ Discotic nematic â†’ Columnar â†’ Crystalline, which have been investigated using differential scanning calorimetry (DSC) accompanied with polarized optical microscopy (POM). Monotropic columnar mesophase was observed during the cooling cycle while discotic nematic mesophase was observed in both the heating and cooling cycles. Density functional theory (DFT) along with IR and Raman spectroscopic techniques were utilized to study the dynamics of molecules during phase transition. To predict the most stable conformation of the molecule, one-dimensional PES scans have been performed along 31 flexible bonds using DFT/B3LYP/6-311G++(d,p) method. Vibrational normal modes were analyzed in detail, taking potential energy contribution into account. The spectral analysis of FT-IR and FT-Raman was done by deconvoluting the structural sensitive bands. The agreement between the calculated IR and Raman spectra and the observed FT-IR and Raman spectra at room temperature confirms our theoretically predicted molecular model of investigated discotic liquid crystal. Moreover, our studies have unraveled the existence of intact intermolecular H-bonding of dimers throughout the phase transitions.

Factors and Radiographic Findings Influencing Patient-Reported Outcomes Following Maisonneuve Fractures.

BACKGROUND: This research adds to the literature by providing prognostic information for physicians and patients regarding the outcomes of operative management of Maisonneuve fractures (MFs). To date, this is the only cohort study of patient-reported outcomes measurement information systems (PROMIS) scores following surgical fixation of MF. Patient outcomes were compared focusing on the mean population with an inter-analysis using basic demographic information, radiographic findings, and patient comorbidities and their respective impact on PROMIS scores. METHODS: A total of 24 patients between 2012 and 2020 met the inclusion criteria and completed PROMIS surveys at a minimum of 18 months postoperatively. Patient charts were reviewed through the electronic medical record (EMR) for demographic information and comorbidities as well as operative variables. PROMIS scores for physical function (PF), pain interference (PI), and depression were obtained via follow-up visits and phone calls. The impact of categorical variables on complications was compared using Chi-Squared tests. Variables were analyzed with a type 3 SS test to stratify independent risk factors' effect on PROMIS scores and to account for confounding variables. RESULTS: PROMIS PF averaged 44.84 and was significantly affected by BMI>30 (p=.033), hypertension (HTN) (p=.026), patients with clinical anxiety or depression (p=.047), and subsequent screw removal (p=.041). PROMIS PI averaged a score of 54.57 and was significantly affected by BMI>30 (p=.0046), coronary artery disease (CAD) (p=.0123), patients with clinical anxiety or depression (p=.0206), and subsequent screw removal (p=.0039). PROMIS depression scores averaged 46.03 and were significantly affected by the presence of CAD (p=.049) and subsequent screw removal (p=.023). CONCLUSION: Patient-reported outcomes following MF surgery demonstrated PROMIS scores within +/- 1 standard deviation of the population-based control, and thus many patients can reasonably expect to return to a level of function comparable to the general population. Nonetheless, the significant effects of patient comorbidities and surgical variables ought to be evaluated and utilized as prognostic indicators when managing patient expectations prior to operative treatment of an MF injury.

SKP2 knockout in Rb1/p53 deficient mouse models of osteosarcoma induces immune infiltration and drives a transcriptional program with a favorable prognosis.

Purpose: Osteosarcoma (OS) is an aggressive bone malignancy with a poor prognosis. One putative proto-oncogene in OS is SKP2, encoding a substrate recognition factor of the SCF E3 ubiquitin ligase. We previously demonstrated that SKP2 knockout in murine OS improved survival and delayed tumorigenesis. Here we aim to define the SKP2 drives transcriptional program and its clinical implication in OS. Experimental Design: We performed RNA-sequencing (RNA-seq) on tumors from a transgenic OS mouse model with conditional Trp53 and Rb1 knockouts in the osteoblast lineage ("DKO": Osx1-Cre;Rb1lox/lox;p53lox/lox) and a triple-knockout model with additional Skp2 germline knockout ("TKO": Osx1-Cre;Rb1lox/lox;p53lox/lox;SKP2-/-). We validated our RNA-seq findings using qPCR and immunohistochemistry. To investigate the clinical implications of our results, we analyzed a human OS patient cohort ("NCI-TARGET OS") with RNA-seq and clinical data. Results: We found large differences in gene expression after SKP2 knockout. Strikingly, we observed increased expression of genes related to immune microenvironment infiltration in TKO tumors. We observed significant increases in signature genes for macrophages and to a lesser extent, T cells, B cells and vascular cells. We also uncovered a set of relevant transcription factors that may mediate the changes. In OS patient cohorts, high expression of genes upregulated in TKO was correlated with favorable overall survival, which was largely explained by the macrophage gene signatures. This relationship was further supported by our finding that SKP2 expression was negatively correlated with macrophage infiltration in the NCI-TARGET OS and the TCGA Sarcoma cohort. Conclusion: Our findings indicate that SKP2 may mediate immune exclusion from the OS tumor microenvironment, suggesting that SKP2 modulation in OS may induce anti-tumor immune activation.

Association and solubility of chlorophenols in CCl4: MIR/NIR spectroscopic and DFT study.

This work provides new information on the effect of position and number of substituents on association and solubility of chlorophenols in CCl4. Using MIR and NIR spectroscopy we examined solutions of 12 chlorophenols at several concentrations. In addition, we calculated (DFT) theoretical spectra and structures of monomers and associates of chlorophenols from dimer to tetramer. The number of substituents at positions 2 and 6 allows to divide studied chlorophenols into three Groups: I (3; 4; 3,4; 3,5), II (2; 2,3; 2,4; 2,5; 2,4,5), and III (2,6; 2,4,6; 2,3,4,5,6). An equilibrium between intermolecular OH⋅⋅⋅OH and intramolecular OH⋅⋅⋅Cl hydrogen bonding depends on position and number of substituents. The extent of association decreases in going from Group I to Group III due to growing steric hindrance near the OH group and the resonance effect from Cl. In chlorophenols of Group I, Cl at positions 3 or 5 weakens the OH⋅⋅⋅OH intermolecular hydrogen bonding, while for Group II it strengthens the OH⋅⋅⋅⋅Cl intramolecular bonding. In contrast, Cl at position 4 has minor effect on association. In the case of Group I, increasing concentration shifts the equilibrium towards solute-solute interactions, whereas for Groups II and III dominate the species with intramolecular OH⋅⋅⋅Cl bonding. The theoretical calculations predict that for monosubstituted chlorophenols of Group I the most stable are non-planar cyclic tetramers, while for disubstituted ones, the non-planar cyclic tetramers and linear trimers have similar binding energies. Chlorophenols of Group II prefer the cyclic non-planar trimers, whereas those of Group III form the planar dimers with an antiparallel orientation of the OH groups. Our study reveals that chlorophenols creating the cyclic associates are better soluble in CCl4 as compared with those forming the linear ones. Hence, one can conclude that in an inert or weakly interacting solvents the solubility is closely related to the structure of the solute associates.