bluuebunny/crossref_metadata_2025 huggingface dataset:
The Biomass and Composition of the Gut Microbiota Modify Anaerobic Metabolism
Authors: Riedl Ruth A , Burnett Colin ML , Pearson Nicole A , Atkinson Samantha N , Ollinger Tomye L , Edwards Robert A , Mokadem Mohamad , Kirby John R , Grobe Justin L
Published: November 2023

The lumen of the gastrointestinal tract is host to a large mass of microbiota, and this environment is largely anaerobic. Previous studies have estimated that bacteria contribute 1–3% (~1–3 kg) of an adult human's body mass and that bacteria typically exhibit an ~7 kcal/kg/hr metabolic rate, which implies a total contribution of 150–450 kcal/d to the host's apparent energy expenditure. Surprisingly, however, few studies have attempted to directly quantify the metabolic rate of the gut microbiota in the in vivo setting. We hypothesized that the biomass and composition of the bacterial community would both contribute to the metabolic rate of the gut microbiota, and that the energy flux through this biomass would be anaerobic in nature. Total resting metabolic rate (tRMR) was assessed using direct calorimetry; aerobic (aRMR) was determined using respirometry, and anaerobic RMR (anRMR) was determined as the difference in results between the two methods. Wildtype C57BL/6J mice were anesthetized with ketamine/xylazine to measure baseline tRMR, and tRMR after surgical removal of the cecum. A subset of animals was also pre‐treated with risperidone. Surgical removal of the cecum, which accounts for ~1% of body mass in mice, resulted in an 8% reduction in tRMR within minutes of surgery (sham n=12 pre 0.116±0.004; cecectomy n=13 pre 0.115±0.004; sham post 0.113±0.004; cecectomy post 0.104±0.005 p<0.05 vs pre, in kcal/hr). Pre‐treatment with risperidone (which we have previously demonstrated to have major modulatory effects upon the composition of the gut microbiome) suppressed tRMR (n=14 pre 0.104±0.004, p<0.05 vs sham pre), and cecectomy in pretreated mice had no additive effect to suppress tRMR (post 0.099±0.004). These results demonstrate that removal of gut microbiota reduces tRMR, subject to the composition of the bacterial community. Next, a cohort of overweight C57BL/6J mice underwent Roux‐en‐Y gastric bypass (RYGB) or sham surgery. After one week, tRMR was significantly increased by 10% in the RYGB group (sham n=8, 0.141±0.003 vs RYGB n=10, 0.154±0.003 kcal/hr, p<0.05) while aRMR remained unchanged (0.140±0.003 vs 0.143±0.003 kcal/hr, p=NS), indicating that the increase in tRMR was completely due to an increase in anRMR (0.001±0.003 vs 0.011±0.003 kcal/hr, p<0.05). These results demonstrate that increased biomass of gut bacteria via RYGB surgery is associated with an increase in energy expenditure, specifically in the form of anRMR. Collectively these data highlight a major contribution of gut microbiota to total energy expenditure in mice, and support the general concept that the energy flux through this biomass is anaerobic and subject to the composition of the bacterial community present.
EXAMINATION OF CHANGES IN INTESTINAL MICROBIOTA INDUCED BY HIGH FAT FEEDING IN MALE ADOLESCENT RATS
Authors: Crawford Melisa Shaunte , Whisner Corrie , Sweazea Karen
Published: November 2023

The gut microbiome has a significant physiological impact in both health and disease. In a healthy individual, microbes influence metabolism, offer protection against pathogens and train the immune system. The dominant bacterial phyla in the human digestive system are gram‐positive Firmicutes and Actinobacteria as well as gram‐negative Bacteroidetes. Recent metagenomic studies show that obesity is associated with reduced Firmicutes and Bacteroidetes while an increase in Actinobacteria is observed. Additionally, the gut microbiome is implicated in the regulation of host metabolic processes, which include producing the short‐chain fatty acids (SCFA), butyrate, acetate, and propionate. However, previous research has shown that prolonged consumption of a Westernized diet can drastically alter gut microbiota and increase the risk of obesity and other metabolic diseases in adults. Although it is known that energy dense foods can alter microbial diversity, less is known about the physiological effects of diet induced changes in the gut microbiome on cardiovascular disease risk in adolescents. Six‐week‐old periadolescent male Sprague Dawley rats were divided into two groups (n=6 each) and fed either a standard rodent chow (18.9% protein, 57.33% carbohydrates, 5% fat) or a high‐fat diet (HFD) (20% protein, 20% carbohydrates [6.8% sucrose], 60% fat) for 6 weeks. Following the diets, fecal pellets were collected and sequenced at the Microbiome Analysis Lab at Arizona State University. Although not significant, the fecal taxonomy analyses suggest changes in the relative abundance of Firmicutes (Chow: 86.3±4.6%; HFD: 80.8±5.0%), Bacteroidetes (Chow: 5.6±2.4%; HFD: 15.0±4.9%), and Actinobacteria (Chow: 0.42±0.17%; HFD: 1.62±0.72%) following the HFD protocol. Understanding how the gut microbiome is altered with poor nutrition may elucidate potential treatment strategies for adolescent overweight and obesity as well as prevention of cardiovascular disease.

Support or Funding Information

Sigma Xi Grant‐in‐Aid Research Program
CD44v9 Emerges in Response to Injury and Contributes to the Regeneration of the Gastric Epithelium
Authors: Bertaux‐Skeirik Nina , Wunderlich Mark , Teal Emma , Mahe Maxime , Sundaram Nambirajan , Gabre Joel , Hawkins Jennifer , Jian Gao , Engevik Amy C , Yang Li , Wang Jiang , Goldenring James R , Qualls Joseph , Helmrath Micheal A , Diwan Tayyab , Mulloy James C , Zavros Yana
Published: November 2023

Cluster‐of‐differentiation gene 44, in particular CD44 variant isoforms (CD44v), emerges during regeneration of the gastric epithelium in response to injury. In particular, CD44v9 is expressed within Spasmolytic Polypeptide/TFF2‐Expressing Metaplasia (SPEM) glands during gastric repair, but the function is unknown. Here we tested the hypothesis that CD44v9 marks a regenerative cell lineage that plays a functional role in gastric repair. Acetic acid injury was induced in CD44‐deficient (CD44KO) and C57BL/6 (BL6) mice. Mouse‐derived gastric organoids expressing CD44v9 were transplanted at the ulcer site of CD44KO mice. Ulcers, expression of CD44v9, proliferation and histology were measured at days 1, 3, 5 and 7 post‐injury. Human‐derived gastric organoids from elderly patients (>55 years) or young patients (14–20 years) were generated and transplanted into the stomachs of NOD scid gamma (NSG) mice post‐injury. Ulcers were induced in NRGS mice expressing human‐derived immune cells (huNRGS) and the immune phenotype analyzed by CyTOF. CD44v9 expression emerged at the ulcer margin during gastric ulcer repair. Compared to BL6 mice that healed within 7 days post‐injury, CD44KO mice exhibited loss of ulcer repair and gastric epithelial regeneration. Transplantation of CD44v9‐expressing gastric organoids into the stomachs of CD44KO mice promoted ulcer repair. Compared to NSG mice exhibiting loss of CD44v9 expression and gastric repair in response to injury, transplantation of human‐derived gastric organoids from young stomachs promoted repair. Transplantation of organoids derived from aged stomachs did not promote repair. Compared to NRGS mice, huNRGS animals exhibited smaller ulcer sizes and an infiltration of human CD163+ macrophages that correlated with an emergence of CD44v9 expression. Thus, CD44v9 marks a regenerative cell lineage. Macrophages infiltrating the injured gastric mucosa may induce the emergence of CD44v9 during regeneration of the epithelium.

Support or Funding Information

This work was supported by NIH 2 R01 DK083402‐06A1 grant, College of Medicine Bridge Funding Program (Zavros), and the University of Cincinnati Graduate School Dean's Fellowship, Albert J. Ryan Fellowship and 2T32GM105526‐04 (Bertaux‐Skeirik).
Genistein is Associated with Increased Colonic Occludin Expression in Female ob/ob mice
Authors: Chompre Gladys , Hernandez Jayna , Cerda Jean , Seguinot Innevy , Hernandez Siomara , Al‐Nakkash Layla , Appleyard Caroline
Published: November 2023

According to the Centers for Disease Control and Prevention statistics from 2014, one third of U.S. citizens are obese increasing the risk of other diseases such as metabolic disorders and cardiovascular diseases. Studies have reported that obesity is characterized by inflammation of the intestinal epithelial barrier. One of its components are the Tight Junction (TJ) proteins including Claudin and Occludin, which mainly function as a physical barrier against diffusion of pathogens, toxins and other antigens. Damage in Claudin‐5 and Occludin is associated with gastrointestinal epithelial inflammation. Interestingly, it has been shown that a diet high in genistein, an estrogen agonist, could have a protective function for the TJ barrier against oxidative stress, bacteria and its products, and inflammatory cytokines. Besides, it can increase vitamin D receptor (VDR) expression in cancer colon cell lines. However, the impact of genistein on gender differences in obesity is not well understood.

AIM

Genistein treatment in female and male leptin‐deficient mice will enhance Claudin‐5, Occludin, and VDR gene expression in proximal colon, whereas IL‐1β gene expression will decrease in both sexes.

METHODS

Female and male leptin knockout mice (ob/ob mice) and wild type littermates (control group) were fed standard rodent chow or genistein‐enriched diet for 4‐weeks. Animals were euthanized and organs were collected including colon. Proximal colon tissue samples were evaluated for microscopic damage and mRNA was extracted for expression of Claudin‐5, Occludin, VDR and IL‐1β.

RESULTS

There was an increase in occludin in ob/ob female mice treated with genistein (p=0.03) compared with control group. Female wild type mice fed genistein had increased VDR, IL‐1β, and microscopic damage (p=0.03) compared with standard chow. Interestingly, VDR and IL‐1β expression was significantly lower in female ob/ob mice fed with genistein compared with the wild type mice fed with genistein (p<0.01). In contrast, male animals did not show any differences in these parameters.

CONCLUSIONS

Our results suggest an association between genistein treatment and enhanced expression of Occludin in ob/ob mice which implies a potential therapeutic effect on permeability function via TJ proteins in the proximal colon in obesity. However, a high genistein diet could have the opposite effect in non‐obese individuals. In summary, genistein supplement in overweight people might provide some beneficial effects.

Support or Funding Information

Supported by NIGMS/INBRE P20 GM103475‐14
Purple Potato Extract Promotes Intestinal Barrier Function and Differentiation by Activating AMP‐activated Protein Kinase
Authors: Sun Xiaofei , Du Min , Navarre Roy , Zhu Mei‐Jun
Published: November 2023

Perturbation of the gut epithelial barrier function induces inflammation and other associated health problems. Purple potato contains a high content of polyphenolic compounds that is known for its beneficial effects. The objective of this study was to evaluate the effect of purple potato extract (PPE) on intestinal barrier function and differentiation, and explore its underlying mechanism in Caco‐2 cells. PPE increased transepithelial electrical resistance and decreased FITC‐dextran paracellular flux in Caco‐2 cells, which were associated with strengthened intestinal epithelial differentiation and tight junction assembly. Furthermore, PPE treatment enhanced AMP‐activated Protein Kinase (AMPK) activity, concomitant with increased expression of CDX2, a key transcriptional factor regulating intestinal epithelial differentiation. Knocking down AMPK using the CRISPR/Cas9 system downregulated the positive effects of PPE on intestinal barrier function and differentiation, in parallel with reduced expression of CDX2. In conclusion, PPE improved gut epithelial differentiation and barrier function via activating AMPK.

Support or Funding Information

Northwest Potato Research Consortium
Complete Bifid Penis in an 84‐Year‐Old Cadaver
Authors: Martin John Richard , Tan Yun , Rana Mohammed Waheed‐Uz‐Zaman
Published: November 2023

Diphallia or penile duplication is a rare congenital variant with an estimate frequency of 1 per 5 million births. The extent of duplication varies widely and typically occurs with other malformations including urogenital, gastrointestinal, and musculoskeletal anomalies. Here, a complete bifid penis was detected incidentally in an 84‐year‐old cadaver in a routine dissection. The phallus was divided longitudinally into right and left halves each having its own glans and prepuce. The urethra was absent in the phallus, opening directly at the base between each phallus. Dissection revealed a corpus spongiosum and a single corpus cavernosum in each phallus. The scrotum was small and contained small sized testes. The spermatic cord took a normal course through the superficial and deep inguinal rings and continued to where the vas deferens ended in well developed seminal vesicles. A small mass of hard tissue was found at the site of the prostate gland but did not have any connection with the vas deferens or seminal vesicle. Examination of other systems revealed no abnormalities. A review of the classification of penile duplication is discussed along with embryologic considerations. Further studies will focus on genetic factors of diphallia.
Persistent Metopic Suture and extra Sacral Foramina: clinically significant anatomical variants
Authors: Gemechu Jickssa M , Geneti Soressa A , Zenebe Abay M , Ewnetu Solomon A , Wirtu Amenu T
Published: November 2023

Lack of knowledge of variability in human morphology and its magnitude compared to normal anatomy may have significant clinical consequences. Accurate knowledge of anatomical variations provides important information in medico‐legal issues, forensic interpretation, diagnostics, imaging and patient management including surgical procedures. It has been hypothesized that a substantial proportion of clinical malpractice is attributed to ignorance of anatomical variations. Therefore, this study aims to assess the variations of metopic suture, sacral foramen and biceps brachii muscle in twenty‐four cadavers used for dissection as part of Gross anatomy course. The study was conducted in five medical schools in Ethiopia in 2015/2016 years. Critical observation, careful dissection and imaging were done accordingly. Our results show a complete persistence of metopic suture in 4.2 % of frontal bone, and the existence of five anterior and posterior sacral foramina in 4.0 % of sacral bone and biceps brachi with three heads of origin in 2.1 % of brachium. Altogether, the findings show the existence of anatomical variants of clinical importance that need special consideration in the diagnosis and treatment of this diversified population of Ethiopia with different socio‐economic background and geographical origin.
The Location of the Rib‐Cartilage Interface‐Importance in Autologous Costal Cartilage Grafting
Authors: Rickards Allen A , Holmes J Scott , Russell Michelle L , Bender‐Heine Adam N , Zdilla Matthew J. , Lambert H Wayne
Published: November 2023

Identification of the rib‐cartilage interface is often done intraoperatively. However, it would be of benefit to understand the locations of the joint lines for pre‐operative planning. Therefore, we conducted a study wherein the rib‐cartilage junctures were identified from 16 cadaveric rib cages. Geometric morphometric analysis was then performed in order to determine a consensus shape of the joint lines. Principle component analysis revealed the most common variations in the locations of the joint lines. The results suggest that the consensus shape, when aligned with the easy‐to‐palpate second costal cartilage, can aid in the identification of joint lines, especially in individuals with little overlying tissue. These results provide a better understanding of the location of joint lines and may aid in the preoperative planning of autologous costal cartilage grafting.

Support or Funding Information

WV Research Challenge Fund [HEPC.dsr.14.13]; WVU Initiation to Research Opportunities (INTRO) Summer Research Program
Rare Bilateral Variation of the Extensor Carpi Radialis Longus Muscle
Authors: Yuan Justin C , Brisson Ryan J , Barremkala Malli , Venuti Judith M , Thompson Brent J
Published: November 2023

Objective

The extensor carpi radialis longus (ECRL) is a superficial extensor muscle with a single muscle belly and tendon. In humans, the ECRL is found on the lateral forearm and serves to extend and abduct the hand at the wrist. While anatomical variations of the musculature of the extensor forearm are uncommon, it is essential for physicians working in this area of the body to be aware of possible variations.

Methods

Dissections of 68 upper limbs from 34 cadavers were performed and the musculature and neurovasculature of the lateral forearm was assessed.

Results

One cadaver displayed an accessory muscle belly bilaterally with a long slender tendon arising from the ECRL muscle and inserting distally near the wrist. Both the accessory muscle on the right upper limb and left upper limb inserted on the abductor pollicis brevis muscle.

Conclusion

This is a rare instance of a bilateral presentation of an accessory ECRL muscle, also known as the extensor carpi radialis accessorius (ECRA). Due to its infrequency, awareness of this accessory muscle and tendon may help physicians avoid improper muscle recognition when visualizing the area. As it is likely that the ECRA only weakly assists with the extension function of the ECRL, when present it may be used as an attractive autograft option for reconstructive surgery with minimal adverse effects.
A Rare Case of Proximal Focal Femoral Deficiency with Overlapping Phenotype of Müllerian Duct Irregularity: An Anatomical and Embryological Anomaly
Authors: Melovitz‐Vasan Cheryl , Kitoh Hiroshi , Vasan Nagaswami
Published: November 2023

Proximal focal femoral deficiency (PFFD) is a rare congenital deformity with various degrees of involvement of the femur with or without other skeletal or organ involvement. Aitken (1969) described and ranked four classes (A‐D) of PFFD severity (least to most severe). In class D the severely shortened femoral shaft appears as an irregularly ossified tuft of bone proximal to the distal femoral epiphysis. During physical examination of a donated body we observed extreme hypoplasia of the left femur with other musculoskeletal anomalies. The donor was a 77‐year old Caucasian female, who died of coronary artery and peripheral vascular disease. Dissection of the cadaver revealed unilateral hypoplasia of the left femur with an aplastic acetabulum, femoral head and patella. Further detailed dissection revealed the absence of the uterus, Fallopian tubes, uterine cervix and the upper part of the vagina. The ovaries were small and atrophied. The external genital structure was normal with blind ending vagina. We also observed involvement of vascular structures. Coexistence of a proximal focal femoral deficiency and congenital absence of the uterus, Fallopian tubes, uterine cervix and vagina is a rare defect (Müllerian duct irregularity) that has not been reported. There is a similarity in the musculoskeletal deformity between the class D phenotype of PFFD we have observed and those with Al‐Awadi‐Raas‐Rothschild syndrome and Fuhrmann syndrome. We report here a rare case of Al‐Awadi‐Raas‐Rothschild syndrome and Fuhrmann syndrome with overlapping phenotype of Mayer‐Rokitansky‐Kuster‐Hauser (MKRH) syndrome typifying congenital absence of the uterus, fallopian tube, cervix and vagina‐Müllerian duct anomaly.

Support or Funding Information

Supported by the department of Biomedical Sciences, CMSRU.
Bilateral Accessory Lung Fissures with a Variant Left Bronchial Tree
Authors: Cohen Samuel , Rajan Benjamin , Zhandosova Annette , Velavan Sumathilatha Sakthi
Published: November 2023

Accessory lung fissures are commonly noted, while bronchial branching abnormalities are less frequently found and are proposed to occur in 0.1–2% of the general population. Multiple accessory fissures, variant normal fissures of both lungs, and an altered left bronchial tree are unique to this case and is being reported due to its clinical significance. During a routine dissection of the thoracic cavity of an 85‐year‐old female cadaver, abnormal fissures were noted on both the right and left lungs, which were photographed. The bronchial branching was further dissected by tracing the bronchus from the hilum to the lung tissue. The right lung had a normal oblique fissure, while the horizontal fissure was shallow and incomplete. The left lung had a normal oblique fissure, and a minor fissure was noted running along the junction of the anterior (S3) and superior lingular (S4) bronchopulmonary segments. Accessory fissures were noted on both right and left lungs. The left bronchus was short and displayed an early division into upper and lower divisions. The dissection of the bronchial tree of the left lung showed trifurcation of the upper division into apicoposterior, anterior, and lingular bronchi. The lower division was normal. The right bronchial tree showed normal division. Examination of the heart revealed an atrial septal defect. The other thoracic and abdominal structures were examined and appeared normal. Knowledge of accessory fissures in the lung have importance in the clinical diagnosis of lobar pneumonia versus atelectasis with imaging and during resection of bronchopulmonary segments. Understanding this bronchial variation and anomalies have implications in diagnosis and endobronchial treatment, including airway stenting, bronchoscopy, endoscopy biopsy, and laser therapy.

Support or Funding Information

Touro College of Osteopathic Medicine, Department of Anatomy
Evaluation of Effect of Pausinystaila yohimbe extract on Liver of Adult Male Wistar Rat
Authors: Okechukwu Hope , Ithentuge Churchill , Ugochukwu Chioma Olejuru , NDUKWE GODWIN
Published: November 2023

Pausinystalia yohimbe is a psychoactive plant that contains the alkaloid yohimbine. It is widely distributed over the counter as an herbal aphrodisiac. Twenty adult male wistar rats weighing between 150g to 200g were used in this research to determine the effects of Pausinystalia yohimbe on the histology of liver. They were divided into five groups; A, B, C, D and E of four rat each. Group A served as control and was fed with feed and water only. Groups B, C, D and E were administered with extract of Pausinystalia yohimbe at a dose of 3mg/kgbw, 5mg/kgbw, 7mg/kgbw and 9mg/kgbw respectively for 21 days. At the end of the experiment, the animals were sacrificed by cervical dislocation after anaesthetized with chloroform. The organ of study (liver) was harvested and sections cut for histological preparation using H and E staining method. Blood for liver enzyme assay was also collected in sterile bottles. Results showed that the level of liver enzyme biomarkers (AST and ALP) were significantly increased at p<0.05 in groups D and E when compared with the control group A while groups B and C were not significantly increased. There was no significant increase in level of ALT in the experimental groups when compared with the control group. Histologically, the results showed that Pausinystalia yohimbe (burantashi) causes moderate to severe destruction of hepatic tissue with severe infiltrate of inflammatory cells, loss of hepatic tissue, congestion of the blood vessel and portal aggregate of inflammatory cells in groups C, D and E. This shows that Pausinystalia yohimbe is capable of causing hepatic damage
Rare Bilateral Morphological Variant of the Renal and Gonadal Blood Vessels. A Case Report.
Authors: Zahoi Delia Elena , Sztika Dorina , Enache Alexandra , Dăescu Ecaterina
Published: November 2023

During the routine dissection of a 68‐year old male cadaver within our Department of Anatomy‐Embryology, a number of anomalies of the renal and gonadal vessels were observed. Each of the kidneys had two renal arteries and two renal veins. On the left side, the superior renal artery gave birth to a superior polar artery and an accessory gonadal artery. The left renal veins formed a ring around the abdominal portion of the aorta. The superior vein (with a preaortic path) had, as tributaries, the first left lumbar vein and an accessory gonadal vein, along with the suprarenal and left gonadal veins. The inferior vein (with a retroaortic path) had two lumbar veins as tributaries. The inferior renal artery and vein were both situated behind the ureter. The left testicle was drained by three veins – one main testicular vein, opening into the left renal vein, and two accessory veins, one also opening into the left renal vein and one directly into the inferior vena cava. On the right side, the two renal arteries ran parallel to each other. The superior artery gave birth to a superior polar artery, while the inferior one gave birth to the right gonadal artery. Both renal veins opened into the inferior vena cava. Awareness of such morphological variants in both renal and gonadal vessels is key to successful surgical interventions and avoiding of associated hemorrhages.

Support or Funding Information

N/A
Natural Antioxidants and Colorectal Cancer: A New Approach
Authors: Jurjus Abdo , Geagea Alice Gerges , Al‐Kattar Sahar , Harmouche Melissa , Harakeh Steve , Diab‐Assaf Mona , Azar Rania , Eid Assad , Massade Liliane , Manfredi Rizzo
Published: November 2023

Introduction

Colorectal cancer (CRC) is a multifactorial disease; genetic epigenetic and environmental factors as well as oxidative stress could be responsible for its onset and progression. Several studies have documented the importance of natural antioxidants in countering oxidative stress and thus preventing colorectal carcinogenesis.

Objective

This study aimed to test the protective effect of a mixture of antioxidants against CRC progression in a colorectal cancer mouse model at cellular, microscopic and macroscopic levels.

Methods

Male C57BL/6 mice were divided into 4 groups, Group 1 given the mixture (M) which contains Curcumin, Resveratrol, Quercetin, EGCG and Cruciferex PB (0.1 mg/mL + DSS (1.5%) + AOM (10mg/kg)). Group 2, M (1 mg/mL + DSS (1.5%) + AOM (10mg/kg)). Group 3, M (1 mg/mL) and Group 4, (DSS (1.5%) + AOM (10mg/kg)). Body weight, intake of drinking water and diet were recorded every day. Two sacrifices were made (week 7 & 14) and biopsies of the liver, small intestine, colon and kidneys were collected and processed for histology and histochemistry.

Results

The mixture (M) improved the clinical signs and symptoms of the animals after the DSS cycle by decreasing the bleeding, diarrhea, inflammation and regaining the weight lost during the past week. Macroscopically, PB was able to decrease the formation of intestinal polyps, vessels vasodilation, inflammation and necrosis of the colon with respect to the controls. Microscopically, PB at both concentrations was able to decrease the infiltration of inflammatory cells, including mast cells and the formation of polyps, compared to the controls.

Conclusion

The mixture (M) seems to have an effect in halting the progression of CRC at the macroscopical and microscopical levels. Further studies are needed to validate its effect at the molecular level.
Significance of Recognizing Anatomical Variations for Medical Education – an Example of the Triceps Brachii Heads
Authors: McDowell Arthur R , Wade Michael D , Ziermann Janine M
Published: November 2023

The triceps brachii muscle is the only muscle in the posterior compartment of the arm comprising three heads. The long head originates from the infraglenoid tubercle of the scapula, the lateral head mostly from the dorsal/posterior surface of the humerus proximal to the radial groove, and the medial head mostly from the dorsal/posterior surface of the humerus distal to the radial groove. It is commonly accepted that all heads of the triceps brachii muscle receive motor innervation by the radial nerve. However, scattered reports of alternative innervations to one or more heads of the triceps brachii can be found. In particular the motor innervation to the long head of the triceps (LHT) seems to be variable as was shown by reports of traumatic injuries to the axillary nerve which were associated with the paralysis of the LHT. This implies that the LHT in those cases was innervated by the axillary nerve. Both, the radial and axillary nerve, are terminal branches of the posterior cord of the brachial plexus. The axillary nerve has an anterior and a posterior branch. The latter emerges at the inferior border of the scapula traversing the quadrangular space, giving off a branch to the teres minor, a branch to the deltoid and a superior lateral cutaneous branch. From a posterior view the radial nerve can be seen through the quadrangular space but this nerve is not traversing the space. Given the spatial correlation of the axillary nerve in the quadrangular space, the insertion of the LHT and the relative distance to the radial nerve in this area, an innervation via the posterior branch of the axillary nerve seems plausible. In our study, we dissected 29 arms in 15 cadavers that revealed radial innervation of all triceps heads, with the radial nerve penetrating the muscles from anterior. This opens the clinically relevant question on how high is the incident of axillary innervated LHTs since the axillary nerve is often damaged (compressed, stretched or even severed) during shoulder dislocation and other injuries. Our results will be relevant for surgical procedures of traumatic nerve injuries as well as for therapeutic treatment (physical and occupational therapies).
Geometric Morphometric Analysis of Costal Cartilage: Considerations for Autograft Selection in Rhinoplasty
Authors: Russell Michelle L , Bender‐Heine Adam N , Rickards Allen A , Holmes J Scott , Armeni Mark A , Lambert H Wayne , Zdilla Matthew J
Published: November 2023

Costal cartilage grafting is commonly utilized in rhinoplasty. Despite the common utilization of costal cartilage autografts, there are differing opinions regarding which costal cartilage levels provide the most ideal grafts. The shapes of costal cartilages are thought to lend themselves to the shapes of certain grafts; however, there has been little study of the shapes of costal cartilages, and most reports have been anecdotal. Therefore, this study aimed to detail the average shapes and most common shape variation of the fifth, sixth, and seventh costal cartilages. A total of 96 cadaveric costal cartilages were analyzed through geometric morphometric techniques. The fifth costal cartilage was determined to have the straightest shape, in general, and would therefore be particularly suitable for nasal dorsum onlay grafting. Likewise, the medial aspects of the sixth and seventh cartilages tend to be straight and are, likewise, suitable for reconstruction of the nasal dorsum. However, surgeons should be aware that the sixth and seventh costal cartilages tend to have an acute curvature in the coronal plane approximately four‐fifths of their total length away from the sternum.

Support or Funding Information

WV Research Challenge Fund [HEPC.dsr.14.13] and

WVU Initiation to Research Opportunities (INTRO) Summer Research program
Accelerometer Assessment of Spinal Joint Crepitus Before and After Spinal Manipulation
Authors: Cramer Gregory D , Budavich Matthew , Bora Preetam , Ross Kim
Published: November 2023

The objective of this IRB‐approved project was to assess spinal joint [facet joint, zygapophyseal (Z) joint] vibrations/sounds (crepitus) during lumbar motion before and after side‐posture spinal manipulation (SM).

Methods

Using methods previously developed to assess joint cavitation, 9 accelerometers were applied to the lumbar region of 5 healthy and 5 low back pain (LBP) subjects, allowing assessment of crepitus. Subjects underwent full lumbar ranges of motion, SM, and repeated ranges of motion, while accelerometer recordings were made. Subjects were also divided into 3 age groups for comparisons (18–25, 26–45, and 46–65 years). Primary outcomes included: incidence of crepitus, crepitus prevalence (average number of crepitus events/subject), crepitus amplitude, and motion(s) producing the most crepitus.

Results

The accelerometry methods were successfully implemented on all subjects. Crepitus was identified as simultaneous deviations from baseline of multiple accelerometers. As in previous work, artifacts had a distinct appearance and were easily distinguished from crepitus. Overall, 23 instances of crepitus were recorded from 15 different Z joints. Crepitus prevalence decreased pre‐ to post‐SM (average pre= 1.4 crepitus/subject, range 0–7 crepitus/subject vs. post= 0.9, range 0–2). Crepitus prevalence progressively increased from the youngest to oldest age groups (pre‐SM= 0.0, 1.67, and 2.0, respectively, range 0–10; and post‐SM= 0.5, 0.83, and 1.5, range 1–5). Prevalence was higher in LBP subjects compared to healthy (pre‐SM‐LBP= 2.0, range 0–7, vs. pre‐SM‐healthy= 0.8, range 0–4; post‐SM‐LBP= 1.0, range 0–2 vs. post‐SM‐healthy= 0.8, range 0–2), even though healthy subjects were older than LBP (40.8 years vs. 27.8 years). Crepitus amplitude was <0.16 mV. Average pre‐SM crepitus amplitude was 0.08 mV, which was greater than post‐SM crepitus, which was 0.03 mV. The pre‐ vs. post‐SM crepitus difference was greater in LBP (pre‐SM 0.09 mV, post‐SM 0.04) than healthy subjects (pre‐SM 0.04 mV, post‐SM 0.03 mV). Flexion produced the most crepitus (pre‐SM eight, 57.1%, post‐SM six, 66.7%, and overall fourteen, 60.9%). For pre‐SM, flexion was followed by right lateral flexion (three, 21.4%) with extension, left lateral flexion and left rotation all producing the same amount of crepitus (one each, 7.1%). Post‐SM flexion was followed by extension (two, 22.2%) and right rotation (one, 11.1%), with no crepitus recorded in the other motions.

Conclusions

Crepitus was greatest in flexion; prevalence increased with age, was higher in LBP than healthy subjects, and overall decreased following SM; crepitus amplitude was lower post‐SM than pre‐SM.

Support or Funding Information

NIH/NCCIH Grant# – 3R01AT000123‐06S2
Metaphysis repair process differ from the diaphysis
Authors: Inoue Satoshi
Published: November 2023

Purpose

In humans, fractures of the long bones mainly occur in the metaphysis such as distal radius. However, most studies of the bone repair process have focused on the diaphysis region, thus little has been reported on the metaphysis repair process. In this study, we compared the bone repair process between the metaphysis and the diaphysis of the mouse tibia.

Materials and Methods

Bone apertures were formed in the tibial metaphysis and diaphysis of mice by 0.8mm drill hole. At indicated times after the surgery, samples were collected, and examined by histomorphometrical (micro‐computed tomography), histological (toluidine blue), histochemical (alkaline phosphatase (ALP)), and immunohistochemical (osteocalcin and type I collagen) analyses. Quantitative real‐time PCR was used to analyze the expression of mRNAs in the periosteum (sox9, type 2 collagen) and bone marrow (osterix, runx2, type 1 collagen).

Results

In the metaphysis, cartilage formation was not detected during the repair period on the periosteum side. The bone aperture was filled with newly formed bone developed from bone marrow at 7 days, and then gradually decreased. In the diaphysis, cartilage was formed around the bone aperture at day 4 and sequentially replaced by bone on the periosteal side. Periosteal callus peaked at day 14, and then remodeled to original architecture. Medullary callus gradually increased to day 14, and the aperture filled with medullary callus transiently at day 14, then medullary callus rapidly resorbed at day 21. In both sites, bone mineral density (BMD) was gradually restored at the bone aperture. At day 14, BMD was higher in the metaphysis than in the diaphysis, but the diaphysis showed significantly high BMD at day 42. In diaphysis, the expression of type 2 collagen mRNA reached to the peak at day 5 following the peak expression of sox9 mRNA reached to the peak at day 4 on the periosteum. However, in the metaphysis, expression of those mRNAs was lower than the diaphysis. Expression of the osteogenic markers in bone marrow, alkaline phosphatase, osteocalcin and type I collagen, appeared earlier in the metaphysis than the diaphysis. In the metaphysis, osterix, runx2 and type 1 collagen mRNAs were markedly upregulated compared with the diaphysis.

Discussion

These results indicate a difference in the bone repair process between the metaphysis and the diaphysis, suggesting a functional heterogeneity of the periosteum and bone marrow mesenchymal cells in response to bone fracture.
The Effects of Bone Morphogenetic Protein‐7 and Interleukin‐1 on Biomechanical Properties of Meniscus Explants
Authors: Taylor Victor Wayne , Hicks Justin , Ferguson Cristin , Danelson Kerry
Published: November 2023

Introduction

The meniscus is a wedge, C‐shape fibrocartilaginous tissue that is essential for the normal biomechanical function of the knee. Damage to the meniscus leads to degenerative changes and loss of function mechanical function of the knee. Cytokines like interleukin‐1 (IL‐1) can inhibit repair process. However, anabolic factors like bone morphogenetic protein‐7 (BMP‐7) may help aid in the repair process. The goal of this project is to observe the effects of BMP‐7 and IL‐1 on the biomechanical properties of the meniscus using microindentation test.

Method

Six‐millimeter porcine meniscus explants were cut using a biopsy punch and cultured in 1% fetal bovine serum media for seven days. Explants were cultured without treatment (control group) or treated with BMP‐7, IL‐1 alpha, and or combined with BMP‐7 and IL‐1 alpha. Microindentation test was used to test the elasticity of the explant tissues.

Results

The elasticity for the explants treated with BMP‐7 (2.480 MPa) were significant compared to explants treated with either IL‐1 alpha (1.501 MPa; p= .001) or both BMP‐7 and IL‐1 alpha (.866; p ≤ .000). This indicates BMP‐7 and IL‐1 have a significant effect on the meniscus biomechanical properties. BMP‐7 increase the elasticity (but not significant) compared to the control explants (2.072 MPa; p= .065), while IL‐1significantly decrease the elasticity (p= .017).

Conclusion

This data shows that BMP‐7 has the potential to exert an anabolic effect on cultured meniscus explants reflected by the presence of increased mechanical compression properties. As a result, the meniscus explant tissues that were cultured with BMP‐7 became more elastic after being cultured for seven days. In addition, meniscus cultured with IL‐1 alpha or a combination of BMP‐7 and IL‐1 alpha became less elastic. Future experiments will continue testing the effect of these two treatments.
Comparative Anatomy of the Anterior Cruciate Ligament Insertions in Primates
Authors: Ruth Aidan , Raghanti Mary Ann , Lovejoy C. Owen
Published: November 2023

Despite a robust body of clinical literature on human entheses, relatively little is known about their anatomy in other species. We sought to document the microstructure of the femoral and tibial insertions of the anterior cruciate ligament (ACL) in a number of mammals, with a particular focus on Primates. Species investigated included those using unguligrade (deer, sheep), digitigrade (lion, tiger), and plantigrade (gorilla, Diana monkey, human) postures. We also explored the relationships between fibrocartilage thickness at ligament insertion sites and body mass.

We found that most anatomical landmarks of the human ACL insertion have homologues in other mammals, including the Lateral Intercondylar Ridge (LIR), the Lateral Intercondylar Posterior Ridge (LIPR), and the Medial Intercondylar Ridge of the Tibia. However, we found that these ridges are not limited termini as is often assumed. Rather, their entheses are expansive and exhibit fibers that extend from both anterior and posterior aspects of these ridges. Considerable variation exists at the lateral border of the tibial ACL insertion, most likely related to spatial distinction of three subdivisions of the ACL (“bundles”) in deer and other ungulates. While primates still display up to three ACL bundles, they adhere more closely to one another and lack distinct septa. Humans share this primate‐typical anatomy.

We measured fibrocartilage thickness of the ACL enthesis on histological thin sections using the method of orthogonal intercepts. Results of a standard linear regression of fibrocartilage thickness and body size indicate that there is no relationship between the natural log of body size as reported in the literature and the natural log of fibrocartilage thickness for either the femoral (F = 1.505, p= 0.1) or tibial (F = 2.57, p = 0.09) insertion sites of the ACL. These data suggest the possibility that, despite the potential biomechanical advantage of substantial fibrocartilage thickness, there may be an upper limit dictated by the (in)ability of nutrients and other cellular signals to diffuse within its extracellular matrix.
New microRNA Biotechnology to Inhibit Inflammation and Regenerate Bone
Authors: Amendt Brad , Cao Huojun , Yu Wenjie , Holton Nathan , Eliason Steven , Hong Liu
Published: November 2023

Current tools for the inhibition of microRNA (miR) function are limited to modified antisense oligonucleotides, sponges, and decoy RNA molecules and none have been used to understand miR function during development. CRISPR/Cas mediated deletion of miR sequences within the genome requires multiple chromosomal deletions to remove all functional miR families due to the duplication of miR seed sequences and family members. We report a novel plasmid‐based miR inhibitor system (PMIS) that inhibits miR family members in cells and mice. The PMIS engineered optimal secondary structure, flanking sequences and specific antisense miR oligonucleotide sequence bind the miR in a stable complex to inhibit miR activity. In cells, one PMIS can effectively inhibit miR family members that share the same seed sequence. A complete family of miRs can be inhibited with a single plasmid. Different PMIS miR inhibitors can be linked together to knockdown multiple miRs expressed from different chromosomes. The PMIS shows no off‐target effects or toxicity and is highly specific for miRs sharing identical seed sequences. PMIS constructs associate with Dicer and Argonaute to form a stable miR inhibitor complex. Transgenic mice expressing PMIS‐miRs reveal different developmental processes affected by miRs. Genome‐wide analyses of PMIS transgenic mice and cells identified new miR regulated gene networks. Complete inhibition of the miR‐17‐92, miR‐106a‐363, miR‐106b‐25, and miR‐200 clusters reveals new mechanisms for bone regeneration and developmental defects for these miRs. These miRs also control inflammation through the direct targeting of pro‐inflammatory cytokines. We report a new tool to dissect the role of miRs in development without genome editing and as a potential new therapeutic reagent.

Support or Funding Information

This work was supported by grants from the State of Iowa Bio Venture group, College of Dentistry and Carver College of Medicine and NIH DE025328 to LH.
Direct Conversion of Oral Epithelial Cells to Progenitor Dental Mesenchymal Cells by a Novel microRNA‐Transcription Factor Pathway
Authors: Mansaray Maurisa N , Sharp Thad , Li Xiao , Huojun Cao , Amendt Brad
Published: November 2023

There are many protocols used for regeneration therapies to regenerate tissues and structures. To date, tooth regeneration practices rely heavily on the use of stem cells, however, these can be difficult to obtain and culture. We have previously introduced a new method of cellular reprogramming by over expression of the transcription factor, Pitx2 and microRNA‐200a‐3p. We now demonstrate that over expression of the Paired Box 9 (Pax9) gene and inhibition of miR‐200a using the Plasmid‐based miRNA Inhibitor System (PMIS‐miR‐200a), converts oral epithelial cells to progenitor dental mesenchyme cells. Transgenic DNA was introduced using the lentivirus packaging system. The microRNA inhibitor system PMIS‐miR‐200a was constructed by NaturemiRI. A two‐staged approach is used to generate stable oral epithelial cells expressing PAX9, cells were sorted and after several weeks were transduced with PMIS‐miR‐200a, to inhibit miR‐200a activity. miR‐200a regulates Wnt and TGFß signaling and inhibition of miR‐200a allows for cells to convert to a progeny phenotype in the presence of PAX9. These cells express early dental mesenchyme and stem cell markers making them suitable for tooth regeneration therapies. Current experiments are using reprogrammed dental epithelial and mesenchyme cells to form a tooth organ. These data demonstrate a new method for cell reprogramming and tissue regeneration.

Support or Funding Information

Support for this research was provided from grant DE013941 and DE018885 from the National Institute of Dental and Craniofacial Research.
Visualizing the attachments and internal architecture of the masseter muscle in Cavia porcellus using contrast enhanced micro‐CT
Authors: Ulm Ashley , Brennan Kevin , Iriarte‐Diaz Jose , Lebowicz Leah A. , Luciano Cristian J. , Vinyard Christopher J. , Druzinsky Robert E.
Published: November 2023

Contrast enhanced micro‐Computed Tomography (CE micro‐CT) renders the anatomy of soft tissues in detail that has, here to for, been unavailable through standard micro‐CT. The CE micro‐CT scans provide a level of contrast in soft tissues that often permits visualization of individual muscle fascicles. Although exquisite details of the internal architecture of muscles can be seen in published slices from CE micro‐CTs, published 3D reconstructions of muscles do not depict those details. The purpose of this project is to utilize CE micro‐CT to segment out the masseter muscles of a specimen of Cavia porcellus, the guinea pig, and visualize the details of the internal architecture of those muscles. The head of an adult guinea pig was soaked in a solution of called iodine potassium iodide (I2KI) for several weeks and then scanned at Northeast Ohio College of Medicine using the Scanco vivaCT micro‐CT. The Materialize Mimics® Innovation Suite will be used to capture this detail in 3D and allow full reconstruction and exploration of the muscle fiber internal micro‐architecture. This data will be put into a 3D PDF to facilitate further research into the attachments, internal structure, and biomechanical function of the masseter and its parts. We hope to develop standardized methods for visualization of muscle architecture that will facilitate future analyses of CE micro‐CT datasets.