bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bioluminescent imaging of hepatic caspase‐3 activity in mice
Authors: Ozaki Michitaka , Haga Sanae , Ozawa Takeaki 
Published: June 2021

Apoptosis‐mediated damage of cells and organs is a serious concern in various pathological situations. Here, we demonstrate caspase‐3 activity by the luciferase‐based optical probe of live liver cells challenged by various stimuli, and also in vivo imaging of caspase‐3 activity of the post‐ischemic liver.METHODSWe developed a novel probe (pcFluc‐DEVD) of cyclic luciferase reflecting caspase‐3 activity. By transfecting an adenovirus cording for pcFluc‐DEVD (AdpcFluc‐DEVD), we investigated whether this probe monitors apoptosis in live AML12 liver cells by staurosporine (STS), Fas‐ligand (FasL) or hypoxia/reoxygenation (H/R), and in the post‐ischemic liver in mice.RESULTSSTS and FasL activate caspase‐3 in caspase‐8‐independent and dependent manner, respectively. First, we measured caspase‐3 activity in STS‐ or FasL‐treated AML12 cells transiently transfected with AdpcFluc‐DEVD. In live AML12 cells, the signals from the probe started to increase immediately and peaked at 2–4 hr after 1 μM STS and 2 μg/ml Jo2 administration, respectively. In both stimuli, caspase‐3 activity was paralleled with apoptotic cell death. Next, we measured caspase‐3 activity in post‐hypoxic AML12 cells by this probe. During hypoxia, caspase‐3 activity dropped to the bottom level. However, it rapidly recovered to the pre‐hypoxic level after reoxygenation and started to rise until 4 – 6 hr post‐H/R.Then, we tried to evaluate in vivo apoptosis by measuring caspase‐3 activity in mouse hepatic ischemia/reperfusion (I/R). 60min of hepatic ischemia caused great increase of capase‐3 activity at 6 – 9 hr post‐I/R, whereas 30 min of liver ischemia caused small increase at 2 hr post‐I/R (Figure).CONCLUSIONSThe caspase3 probe successfully illustrated apoptotic damage in vitro and in vivo. Visualizing apoptotic damage of cells/organs may provide important insights into various physio‐pathological conditions.
pH Regulatory Transporters in Pancreatic Ductal Adenocarcinoma (PDAC)
Authors: Kong Su Chii , Christensen Sandra , Hoffmann Else Kay , Novak Ivana , Pedersen Stine Falsig 
Published: June 2021

PDAC is among the most aggressive human malignacies, with a 5‐ year survival rate of 1–4%. Elucidation of specific molecular alterations in PDAC is urgently needed to devise new treatment strategies. Malignant tumors with rapid progression such as PDAC often undergo extensive metabolic changes resulting in increased acid production. We and others have shown in other cancers that this leads to the upregulation of acid extruding transporters, which in turn favors further cancer progression. The aim of our studies is to elucidate the regulation and roles of pH regulatory ion transporters in PDAC. We used qPCR to screen a panel of 4 human PDAC cell lines (AsPC‐1, BxPC‐3, Panc‐1, MIAPaCa‐2) against a normal human pancreatic ductal cell line (HPDE). The Na+/H+ exchanger 2 (NHE2) was upregulated in 3 of 4 cell lines, whereas NHE1 was upregulated only in BxPC3 cells; the V‐type H+‐ATPase subunit a3 was upregulated in all PDAC cell lines; all cell lines except BxPC3 exhibited upregulation of either AE2 or AE3 Cl−/HCO3−exchangers, and also the Na+‐HCO3− transporters SLC4A7 and –A8 tended to be upregulated in PDAC; surprisingly, the H+/lactate cotransporter MCT1 was downregulated in all PDAC cell lines and a similar trend was seen for MCT4. In conclusion, PDAC cell lines exhibit dramatic changes in the mRNA expression level of pH‐regulatory transporters. Ongoing experiments are addressing the functional relevance of this in PDAC.Funding: Marie Curie Actions (ITN IonTraC)
A novel plate reader based assay for inhibitor studies of slc4a10 protein and other acid/base transporters
Authors: Nguyen An Tuyet , Praetorius Jeppe , Damkier Helle Hasager 
Published: June 2021

The function of members of the slc4a family of HCO3− transporters is commonly determined using fluorescence‐microscopy of tissue loaded with pHi sensitive dyes. The cells are subjected to an acidification induced by NH4Cl and the Na+ dependent pHi recovery is determined in the presence of CO2/HCO3−.The purpose of the present study was to implement the method in a plate‐reader based assay to increase the throughput of the method by measuring an entire cell population, and performing multiple experiments in parallel, thus improving stability and reliability of data.A 3T3 NIH‐fibroblast cell line was stably transfected with the Na+/HCO3− transporter encoded by slc4a10. pHi of cells was measured in 8 wells of 96‐well plates in an Enspire® platereader using the fluorescent pHi indicator BCECF. Control experiments included DIDS, an inhibitor of bicarbonate transporters.The Na+ dependent pHi recovery in slc4a10 transfected cells was 17.8±5.35 ×10−4 pH units/s (n=12). 200 μM DIDS inhibited the recovery by 65% (6.16±3.35 ×10−4 pH units/s, n=12). The endogenous Na+/H+ exchange in the cells was similarly reduced to 66% by 10 μM EIPA in the absence of CO2/HCO3−.In conclusion, this method is a novel and valuable tool for high throughput screening of pharmacological agents inhibiting the acid/base transporters.
Thick ascending limb‐specific nuclear receptor ERRβ regulates renal ion transports
Authors: Crambert Gilles 
Published: June 2021

Renal functions are regulated to cope with the demands of the organism through activation of different receptors. Among them, the family of the nuclear receptors (NR) participates at the long‐term processes. Identity of the NR expressed in the kidney is known, but their quantitative localization in this ultrastructured organ remained poorly described. Using qPCR on microdissected mouse renal nephronic segments, we established a quantitative expression map of NR along the nephron. This map can serve to identify NR with specific localization. Thus, we unexpectedly found that the estrogen‐related receptor β (ERRβ) is expressed predominantly in the Henle loop (TAL). In vivo treatment with an ERR inverse agonist (diethylstilbestrol) showed a link between this receptor family and the expression of NKCC2, and resulted in phenotype presenting some similarities with the Bartter syndrom (hypokalemia, urinary Na+ loss and volume contraction). We then used a mouse TAL cell line to dissect the mechanisms by which NKCC2 and other transporters are modulated after stimulation of ERRβ with a selective agonist (GSK4716) using CHiP experiments.The TAL segments being involved in the regulation of extracellular volume, blood pressure, Ca2+ and Mg2+ homeostasis, identification of ERRβ as a new regulator of ion transport in this particular segments may contribute to better understand dysfunctions of these parameters.
TRAINING 3‐YEAR STUDENT ON EVIDENCE‐BASED MEDICINE
Authors: Tuleshova Gulnara , Abduldayeva Aigul , Derbissalina Gulmira , Delellis Nailya , Êàrp Leonid 
Published: June 2021

Modernization of medical education in the Republic of Kazakhstan in the light of the Bologna process envisages the need for staff training of the new formation. In this regard, the ongoing reform of medical education in Kazakhstan focuses on the quality of professional graduates training of medical schools. In 2006, a new national state standard of medical education of the Republic of Kazakhstan was worked out, where the final goals of specialist training in the public health service at diverse education stages are clearly marked. The peculiarities of SES‐2006 are: the transition to 3‐level education, a focus on competence‐orient education, availability of components for the selection and the emergence of new disciplines, including such disciplines as “Evidence‐based medicine”, “Communication skills”, “Biostatistics” “Epidemiology”. The Knowledge of the issues of evidence ‐ based medicine at the present is necessary for each clinician. Medical students in training must learn to think and work in accordance with the principles of EBM.The Purpose of training on “Basics of EBM” is the creation of knowledge bases of EM, the formation of skills that will permit to evaluate medical information critically for rational use in further practice. The objectives of the training are to acquaint students with the basic terms of EM, the main types of clinical research, usingmethods of EM database, to develop the ability.
Training Efficient Nutrition Counseling in Virtual Reality
Authors: Kohlmeier Martin , Adams Kelly , Morris Evan , Powell Margo 
Published: June 2021

Healthcare providers are tasked with providing nutrition guidance to their patients, something they are neither taught adequately nor given an opportunity to improve their counseling skill under supervision. Unmentored learning on the job is unlikely to help them conduct effective and efficient sessions. We have created an online virtual reality (VR) environment where learners can practice how to counsel patients about lifestyle change that lowers their blood cholesterol levels.Seamless patient video responds to user prompts. Input items support selection from a range of domains or sub domains, thus allowing the user to freely navigate through content space just like in a natural dialog. The patient responses depend both on the selected topic and the specific phrasing of the user's input. Diverse patient scenarios with differences in stage of change and attitude allow multiple sessions without repetition of the dialogs. The user can select a training mode of the application and get continuous visual feedback from a dashboard. The gauges indicate accuracy of the user's nutrition guidance, consideration of the patient's stage of change, and reliance on motivational interviewing techniques. The sessions are timed to emphasize the importance of efficient interactions. After each counseling session, the application shows a dialog transcript linked to a display of the continuously recorded performance metrics.
Obstruction of the thoracic duct(s) causes chronic lymphedema of the upper limb in patients presenting with symptoms of thoracic outlet syndrome: MRI/MRA/MRV
Authors: Collins James Douglas , Saxton Ernestina , Gelabert Hugh , Ahn Samuel , Carnes Alfred Edward , Daniels Portia 
Published: June 2021

INTRODUCTIONObstruction of the thoracic duct(s) causes chronic upper extremity lymphedema. Lymphatics have bicuspid valves like the venous system. Metaplastic fibrosis resulting from obstruction of lymph drainage in the upper extremities impedes vertebral venous plexus / Batson's plexus circulation. The upper limb undergoes painful swelling into the axilla and compresses the brachial plexus. Patients present with tingling/numbness and weakness of the affected arm; lightheadedness; blurred vision and floaters in the visual fields, and tinnitus with whooshing sounds in the ear on the affected side and increased hair growth and nail changes in the affected limb.METHODSBilateral MRI/MRA/MRV displays the sites of obstruction of lymphatics, draining veins of the neck and the subclavian and axillary arteries with binding nerve roots. Monitored multiplanar images are acquired on a 1.5 Tesla GE Signa LX unit, 44 cm field of view, 512 × 256 matrix and saline water bags to enhance signal to noise ratio and Fast Spin Echo (FSE) to display lymphedema (JNMA 1999; 91:333–341).RESULTSTwo patients were selected: one with acquired obstruction of the thoracic lymph duct with lipoma extending into the left hemithorax, the other with developmental fibrosis of the right thoracic duct complicated by pulmonary emboli. Cannulation of the lymphatics could not be performed because of obstructed lymphatic flow.CONCLUSIONSMRI/MRA/MRV is the only alternative to lymphangiogram in patients with obstruction to lymph flow.
Genetic analysis of dermal bone fragmentation in a natural model system.
Authors: Gross Joshua B. , Krutzler Amanda J. , Bruns Lauren E. 
Published: June 2021

Natural model systems that reside in extreme environments can provide powerful insights into fundamental biological processes. For instance, many of the genetic and cellular regulators of normal dermal bone formation have been clarified. However, the precise genetic and molecular bases for altered bone phenotypes evolving under environmental pressure remain largely uncharacterized. In this study, we evaluated a dermal bone fragmentation phenotype that occurs in the blind Mexican cavefish (Astyanax mexicanus). We investigated this phenotype using exhaustive cranial bone measurements in the context of an F2 pedigree of surface fish × cavefish hybrids. We determined that fragmentation of the suborbital bones occurs in a largely asymmetric fashion. Further, we clarified the genetic architecture for left‐sided and right‐sided suborbital bone fragmentation from our hybrid pedigree. Our results indicated that while the area of the face occupied by suborbital bones is constrained between different‐sized individuals, the pattern and severity of fragmentation could not easily be predicted by co‐occurrence of other degenerative characteristics. Our results may indicate a pleiotropic interaction between genes selected in the cave environment for constructive traits that interfere with normal patterns of bone ossification. Research supported by NIDCR (NIH) R03DE022403.Grant Funding Source: grossja@ucmail.uc.edu
Temporomandibular joint articular disc: Providing a structural scaffold for use in tissue bioengineering of replacement constructs
Authors: ten Hove Steven Holterman , Wilson Tim , Shimizu Michael , Merrifield Peter , Galil Khadry 
Published: June 2021

The temporomandibular joint's (TMJ) articular disc facilitates normal function in the healthy joint. Temporomandibular disorders (TMDs) are associated with disc derangement or damage resulting in joint dysfunction. TMD can progress to an end‐stage where surgical removal of condyle and disc are performed. No adequate prosthetic replacement exists for the disc. A dimensional model for the disc is yet to be provided. Our objective is to assess the dimensions of the articular disc providing a structural scaffold for tissue engineering of replacement constructs. Discs were dissected bilaterally from 12 cadavers (56–100yrs). Linear surface dimensions of the discs were measured using digital calipers to assess absolute and regional anteroposterior width (APW) and mediolateral length (MLL). Regional measurements were taken perpendicularly at relative ratios of the absolute dimensions (50% central; 20% from borders). All measurements (mm) were repeated twice by 2 observers to assess reliability. Mean absolute APW (15.31±1.08) and MLL (24.71±2.29) were calculated from 9 specimens along with mean values for 6 regional surface measurements and 9 regional depth measurements per disc. Within the limits of this study it is possible to provide reliable dimensions for use in creating a structural scaffold of the TMJ articular disc.Grant Funding Source: Departmental Funding
Enthesophyte on Radius
Authors: Singh Rajani 
Published: June 2021

During examination of radii in the osteology lab of KGMU three (two radii of left hands and one of right hand ) out of assorted one hundred radii were having longitudinal and transverse enthesophytes. The lengths, heights and thicknesses of these longitudinal enthesophytes in the left two radii were 2.2/0.7/0.2 cm, 1.9/0.7/0.4 cm and 2.2/0.6/0.4 cm in one right radius. The lengths, heights and thicknesses of those transverse enthesophytes in the left two radii were 1.1/0.3/0.2 cm, 0.8/0.2/0.2 cm and 0.7/0.1/0.1 cm in one right radius. This extra bony growth may impinge on the surrounding structures leading to neurovascular complications and restriction of biomechanical movements of forearm. The study will be of paramount importance to anatomists, radiologists and clinicians. The defect is virgin and seen for the first time so the case has been reported.No external support is available to the author
Vagus Nerve's Topography in the Carotid Sheath: preliminary results
Authors: Yendo Tatiana Mina , Docko Ana Jung , Hojaj Flávio Carneiro , Akamatsu Flávia Emi , Andrade Mauro , Jacomo Alfredo Luiz 
Published: June 2021

The knowledge of the vagus nerve's location within the carotid sheath is essencial in many surgical contexts, such as thyroidectomy. Intraoperative neuromonitoring may prevent iatrogenic nerve injury. There are only few studies about vagus nerve and its topography in the carotid sheath. This study aims to analyze the variation of the vagus nerve topography and its relation with the common carotid artery and the internal jugular vein, at the infrahyoid region.We dissected 10 carotid sheaths from 5 cadavers and the position of the vagus nerve and its relationship with the common carotid artery and the internal jugular vein were determined. Based on the carotid sheath cross‐section, the location of the vagus nerve was classified in four quadrants: between the common carotid artery and the internal jugular vein, but anterior to the carotid artery's cross section (A), between the common carotid artery and the internal jugular vein, but posterior to the carotid artery's cross‐section (B), posterior and medial to the carotid artery's cross section (C) and posterior and lateral to the internal jugular vein's cross‐section (D).We found 6 vagus nerve in position (A), 1 in position (B), 0 in position (C) and 3 in position (D).The knowledge of variations and their prevalence regarding the position of the vagus nerve as described in this preliminary study may contribute to avoid iatrogenic lesions in cervical surgeries.
CUTANEOUS RECEPTORS OF THE EPITHELIUM OF LABIA MINORA IN PREPUBERTAL GIRLS
Authors: Schober Justine M , Alguacil Nieves Martin , Pfaff Donald , Mayaglou Lazarus , Aardsma Nathan 
Published: June 2021

IntroductionSparse information is available regarding terminal sensory nerve endings within the glabrous skin of the female external genitalia. Receptors sense tactile stimuli encoding stimulus position, changes in temperature, pressure and pain. Diversity of possible sensations suggests the presence of a variety of receptor types. This neuro‐histological study seeks to document presence and characteristics of cutaneous sensory receptors in female genital tissue.Materials and MethodsLabial skin samples from ten girls who underwent surgery for labial fusion were waste tissue, obtained after informed consent from parent or legal guardian. Cajal‐type silver impregnation, immunocytochemistry against protein gene product (PGP) 9.5, and neuron‐specific enolase (NSE) was performed.ResultsDense innervation of labia minora epidermis with intense staining along the introital versus the external border was noted. Free nerve endings were identified in papillary dermis. Sensory corpuscles: non‐capsulated, Meissner‐like corpuscles, scattered in dermal papillae interdigitated with epidermal ridges of the skin, encapsulated corpuscles protruded into epidermis from deep dermis, Merkel cell‐neurite complex was identified in stratum basale and spinosum. Encapsulated corpuscles and cells located in the inner and outer core were strongly positive to PGP 9.5.
Vascularization of the Pes Anserinus Superficialis
Authors: Hirtler Lena , Faber Mike , Weninger Patrick , Streicher Johannes 
Published: June 2021

IntroductionThe aim of this study was to evaluate the vascularization of the superficial pes anserinus and its inserting tendons with regard to its potential use as a vascularized graft for the reconstruction of the anterior cruciate ligament.MethodsTwelve fresh cadaveric knees were used in this study. The inferior medial genicular artery was identified at its origin from the popliteal artery and incised to be perfused with a methylene blue solution. Its intra‐ and peritendinous course was evaluated macroscopically and histologically.ResultsThis study proved that the superficial pes anserinus is a highly vascularized structure. The vascularization is potentially able to supply the inserting tendons of the semitendinous and gracilis muscles retrogradely.ConclusionThe results of this study indicate that the option of a pediculated cruciate ligament reconstruction with both tendons of the semitendinous and gracilis muscles inserting at the superficial pes anserinus is theoretically possible. A pediculated reconstruction of the anterior cruciate ligament could decrease postoperative morbidity of the transplant and therefore enshorten the vulnerable healing period after surgery.Grant Funding Source: N/A
The Endoplasmic Reticulum Stress and Unfolded Protein Responses in Sulfur Mustard induced Mouse Skin Wound Repair
Authors: Chang Yoke‐chen , Wang James D , Gordon Marion K , Gerecke Donald R 
Published: June 2021

The endoplasmic reticulum stress (ERS) pathways activate the unfolded protein response (UPR) to mediate cell survival against severe stress in an attempt to prevent ERS mediated apoptosis. Sulfur mustard (SM), a chemical warfare vesicating agent, causes significant delayed wound repair in skin. This study explored the contribution of UPR and ERS to SM cutaneous injury in mice. We performed time course studies using the mouse ear vesicant model and showed histopathology changes over time. RT‐PCR demonstrated at least a 2‐fold increased of the ER stress survival gene, GRP78; UPR gene, ATF6; and proapoptotic gene, CHOP as time progressed from 24h to 168h post‐SM exposure. Western blot analysis demonstrated increased GRP78, CHOP, and full length and cleaved (activated) forms of ATF6 as time progressed. This was confirmed by immunofluorescence which showed significant nuclear staining of cleaved ATF6 in the hyperplastic epidermis at 168h post exposure. Double immunofluorescent labeling for GRP78 and CHOP at 72h post exposure showed abundant cytoplasmic staining of GRP78 in the basal, proliferating epidermis. In contrast, the expression of CHOP was more numerous in the migrating keratinocytes. This suggests that the migrating cells are targeted for apoptosis while the basal, proliferating cells are targeted for survival. The continued proliferation of the basal cells and their subsequent conversion to migrating cells may in part explain the delayed wound repair frequently observed in SM skin injury.Grant Funding Source: NIH grants AR055073, EY09056, and ES005022.
Ontogeny of Pan troglodytes and Macaca mulatta Metatarsal Cross‐sectional Properties and Comparisons to the Miocene Catarrhine Proconsul
Authors: Organ Jason M. , Dunsworth Holly M 
Published: June 2021

Catarrhines differ in foot use, function, and anatomy between species and within species through ontogeny. For example, adult Pan and Macaca differ in hallucial anatomy and also in the relative distribution of ground reaction forces. Further, they experience different growth in hindlimb proportions, which may correlate with differences in locomotor ontogeny. These differences should be reflected in metatarsal (MT) cross‐sectional geometries (CSG). To test this hypothesis we measured the CSG of MTs 1–5 in a cross‐sectional ontogenetic sample of 80 M. mulatta and 74 P.troglodytes. Most of the CSG were collected with biplanar radiography and the latex cast method (LCM). However, an additional 28 juvenile M. mulatta individuals, too small for LCM, were recently scanned with microCT. Midshaft CSG were calculated using ImageJ. As expected, CSG are significantly different in both absolute and relative magnitude and patterning across the rays between the two species as well as the age groups within them, particularly in the hallux where Pan shows greater bending and torsional resistance. These results provide insight on foot use in fossils. MTs from six Proconsul individuals ranging from infant to adult were scanned with microCT. Proconsul MT CSG are similar in magnitude to those of Macaca, but share relative hallucial patterning with Pan. This research was funded by the Leakey Foundation and Penn State University.Grant Funding Source: N/A