bluuebunny/crossref_metadata_2025_split huggingface dataset:
Menstrual cycle change in thalamic glutamate decarboxylase and GABA in response to TMJ pain in rats (726.4)
Authors: Umorin Mikhail , Bellinger Larry , Kramer Phillip 
Published: June 2021

Women report TMJ pain more often than men and epidemiological evidence suggests the pain response varies between men and women and over the menstrual cycle. Recent studies show TMJ responsive neurons projecting to the thalamus have different activities in male and females. Using a rat model our lab performed whole genome array screens on thalamic tissue of cycling rats. The results indicate that the transcript of both glutamate decarboxylase 1 and 2 (GAD67 and GAD65, respectively) are significantly up‐regulated in the thalamus of proestrus (high estrogen, progesterone and luteinizing hormone concentration) versus diestrus/metestrus (low estrogen, progesterone and luteinizing hormone concentration) rats. On the other hand, GABA expression (by ELISA assay) did not significantly change in the thalamic tissue during the proestrus phase. The differences in response of the enzyme and its product could be explained by the distribution of each enzyme within individual neurons. In conclusion, sex steroids could alter thalamic GABAergic signaling to effect TMJ hyperalgesia through regulation of GAD.Grant Funding Source: Supported by NIH R01 NIDCR grant: DE022129 Estrogen and TMJ Pain
An anatomical update of the aortic plexus with clinical implications for retroperitoneal surgery (726.6)
Authors: Beveridge Tyler , Johnson Marjorie , Power Adam , Power Nicholas , Allman Brian 
Published: June 2021

The aortic plexus is a sympathetic network of pre‐ and post‐ganglionic nerves overlying the abdominal aorta. The literature minimally describes the anatomy of this plexus and little research has been completed to anatomically define the nerves of this plexus. This study aimed to anatomically map the aortic plexus through dissection of fresh, frozen human cadavers (n=7). Using standard histochemical procedures, ganglia of the aortic plexus were verified with H&amp;E staining and anti‐tyrosine hydroxylase staining confirmed the presence of sympathetic neurons. All specimens (100%) had 4 distinct ganglia that were supplied by right and left, L1 and L2 lumbar splanchnic nerves; 3 documented ganglia (right/left spermatic ganglia and inferior mesenteric ganglion), and 1 previously‐unidentified ganglion. Moreover, the prevalence of the accessory splanchnic nerves (right/left accessory L1 lumbar splanchnic 29%/29%; right/left accessory L2 lumbar splanchnic 14%/86%) and pre‐aortic nerves (inferior branch of right spermatic 86%; right/left intermesenteric nerve 29%/71%) were documented in each specimen to assess variability in the plexus. Clinically, our results could have implications for preserving fertility in men and sympathetic function to the hindgut and pelvis during retroperitoneal surgeries.Grant Funding Source: Supported by Canadian Urology Association Scholarship Foundation Grant
Immunohistochemical staining for neurotransmitter‐related antigens in nerves fixed with high concentrations of glutaraldehyde (726.7)
Authors: Carvalho Carolina , Castania Jaci , Salgado Helio , Fazan Valéria Paula , Llewellyn‐Smith Ida 
Published: June 2021

Immunohistochemistry (IHC) demonstrates specific antigens within tissues or cells and is usually done on formaldehyde‐fixed tissues. However, for a detailed quantification of axons within nerve bundles, glutaraldehyde fixation is mandatory. We aimed to develop an IHC technique for nerves fixed with high concentrations of glutaraldehyde. Phrenic nerves from 8 male Wistar rats were removed, fixed in phosphate‐buffered 2.5% glutaraldehyde and cryoprotected in 30% sucrose. Slide‐mounted cryostat sections (16 µm thick) were incubated for 24 hr in goat polyclonal anti‐choline acetyltransferase (ChAT; 1:1,000) or rabbit polyclonal anti‐substance P (SP; 1:10,000) then transferred to biotinylated donkey anti‐goat or donkey anti‐rabbit immunoglobulin (1:500; 24 hr) before immersion in Extravidin‐HRP (1:1.500; 24 hr). Immunoreactivity in axons was revealed with a nickel‐intensified diaminobenzidine reaction. This method reproducibly revealed individual axons that contained immunoreactivity for ChAT or SP within the phrenic nerve. These results show that peptides and large proteins can be detected by IHC in nerves that have been immersion‐fixed in a high concentration of glutaraldehyde. This method is reliable for identifying neurotransmitter‐related antigens and will allow quantification of axons with specific physiological roles, which can be inferred from their neurochemical phenotypes.Grant Funding Source: FAPESP, CAPES, CNPq and FAEPA
Examining the function of TPX2 during neurite morphogenesis (727.2)
Authors: Hwang Eric , Chen Wen‐Shin , Chen Yi‐Ru , Huang Yung‐An 
Published: June 2021

A functional nervous system depends on the intricate connections between neurons. To establish appropriate connections between neurons, the morphogenetic process of the neuron must be carefully regulated. Neuritogenesis is the morphogenetic process that regulates neurite extension and connection. It has been shown that the microtubule cytoskeleton and microtubule‐associated proteins are indispensable for this process. The microtubule‐associated protein, targeting protein for Xklp2 (TPX2) plays critical roles during mitosis. TPX2 is required to generate a stable bipolar spindle. It localizes to the nucleus during interphase and to the spindle poles during mitosis. We previously identified TPX2 as one of the most abundant proteins on neuronal microtubules, suggesting that TPX2 may play a role during neuritogensis. To examine the function of TPX2 in neurons, we depleted TPX2 in mouse primary hippocampal neurons using shRNA and observed a decrease in neurite length and branching number, suggesting TPX2 is involved in the neurite elongation and branching process. To determine the mechanistic basis of TPX2 on these processes, we examined the localization of TPX2 in hippocampal neurons. We observed that TPX2 localized primarily to the centrosome and interacted with gamma‐tubulin. Additionally, TPX2 also distributed in the neurites and bound to the microtubule cytoskeleton. To determine how TPX2 affects the microtubule cytoskeleton, we analyzed the dynamics of microtubules in TPX2‐depleted neurons by examining the microtubule plus‐end binding protein EB3. However, we did not observe any change in microtubule plus‐end dynamics or microtubule emanating frequency in TPX2‐depleted neurons. These results indicate that TPX2 participates in the neuritogenesis process and may exert its effect via neurite‐localized and microtubule‐bound population.Grant Funding Source: Supported by National Science Council of Taiwan
Vitamin D3, testosterone, epigenetics, and nocioception: an evolving concept of neurohomeostasis (727.3)
Authors: Seigel Eric , Thomas Marc , Tulp Orien , Einstein George 
Published: June 2021

To determine the effects of hormonal contributions to neurosignaling and clinical pain management, a study was undertaken to correlate the effects of plasma concentrations of vitamin D3, Testosterone, markers of epigenetics, and nocioception on outcomes via meta analysis of existing studies. Vitamin D3 and Testosterone have long been shown to be linked to molecular expression of numerous gene products, including calbindin, the homer‐1 gene, cytochrome P24, cytochrome P27, Bone Morphogenetic Proteins [BMPs] and others, all of which are proposed to have supporting molecular roles in neurophysiologic processes and in nocioception. The clinical descriptions that directly link Vitamin D3 or testosterone to pain are inconsistently linked, however, but the composite effects of relevant studies support an emerging concept of nocioceptive homeostasis and neuroplasticity that is in part epigenitcally mediated emerge, and thus include a likely contributory role for steroid hormones in the process of nocioception and certain clinical elements of pain management.Grant Funding Source: Institutional Resources
Myelinated fiber distribution in the aortic depressor nerve of male and female spontaneously hypertensive rats (728.22)
Authors: Amorim Milena , Castania Jaci , Salgado Helio , Fazan Valéria Paula 
Published: June 2021

Aortic depressor nerve (ADN) is an afferent nerve that contains myelinated (fast conducting) and unmyelinated (slow conduction) fibers that send information from aortic mechanoreceptors to central nervous system [1]. Previous studies form our laboratory [2,3] in spontaneously hypertensive rats (SHR) have demonstrated morphological alterations on the ADN in SHR, using male and female animals indistinctively. In this study we investigated the ADN fiber distribution comparing male and female adult (20 weeks old) SHR. After the (AP) and heart rate (HR) recordings, together with the ADN spontaneous activity, the left ADN were prepared for light microscopy and morphometry. Myelinated fiber distributions were compared between proximal and distal segments in the same gender and between genders. Myelinated axon and G ratio distributions were also investigated. Myelinated fiber diameter range from 1 to 5 µm, with peak (~40% of fibers) at 2 µm while myelinated axon size range from 0.5 to 4 µm, with peak (~50% of axons) at 1.5 µm. No differences were observed between segments or genders. This indicates that gender does not influence on the ADN myelinated fiber size, allowing the use of male and females for morphological studies.[1] Krieger and Marseillan, Am J Physiol, 1963:205; [2] Fazan et. al, J Auton Nerv Syst 77, 1999:133; [3] Fazan et. al, Am J Physiol Heart Circ Physiol 280,2001:H1560.Grant Funding Source: FAPESP, CAPES, CNPq and FAEPA
Structural, morphologic and morphometric study of the phrenic nerve: association between experimental diabetes and arterial hypertension (728.24)
Authors: Ferreira Renata , Rodrigues Anaceres , Castania Jaci , Salgado Helio , Fazan Valéria Paula 
Published: June 2021

Although the association between hypertension and diabetes is common in clinical practice, it is not well known the contribution of each of these diseases in the development of diabetic neuropathy in patients with hypertension. We investigated the presence of peripheral neuropathy, through morphology and morphometry analysis of the phrenic nerve in normotensive rats (Wistar) and spontaneously hypertensive rats (SHR) with experimentally induced chronic diabetes. Diabetic animals received a single injection of streptozotocin (STZ) intravenously while controls received vehicle on the same date. Twelve weeks after injection, arterial pressure and heart rate were recorded and the right and left phrenic nerves were removed and prepared for embedding in epoxy resin and computer morphometry. The phrenic nerves of SHR and diabetic SHR (SHR + STZ) showed reduced number and density of myelinated fibers and Schwann cell nuclei, compared to Wistar. The SHR and SHR+STZ rats showed myelinated fiber size distributions skewed to the right, with an important reduction of the small myelinated fibers. This finding was more pronounced on diabetic SHR when compared to normotensive diabetic animals. We suggest that hypertension causes a reduction of small myelinated fibers and the association of the hypertension and diabetes did not interfere with the small fiber neuropathy present in the hypertensive animals.Grant Funding Source: CAPES, FAPESP, CNPq and FAEPA
3D printing from MRI data of stroke and Alzheimer’s disease subjects: an educational model of neurologic disease (728.4)
Authors: Gardiner Brett , Wisco Jon 
Published: June 2021

Normal human anatomy used in the classroom is not reflective of variations confronted in pathology subjects. Current commercialized models are not products of real data, rather representations of it. While learning complicated medical anatomy, students take an enormous leap from the anatomy lab to situational surgical settings. 3D models can bridge this gap in medical education without patient risk, particularly for the brain where surface regions have strong associations to specific physiological activity. Subject specific models are especially advantageous for comprehending real surface morphology of neurologic diseases. Using rapid prototype (RP) technology, we have developed an accessible process to produce physical 3D models from specific MRI data of stroke and Alzheimer’s subjects. The neuroanatomical abnormalities modeled from real data by our 3D printouts will educate students on the anatomical variations encountered in an authentic clinical scenario of neuropathology. Our methods consisted of three phases: (1) image acquisition, (2) post‐processing imaging data with segmentation, and (3) 3D printing. By delineating cortical regions we have provided a unique multidimensional facet of clinically accurate data not yet available to the classroom. This powerful and versatile technique allows students and professionals to visualize the inherently complicated structures as seen in clinical neuropathology. From students in the classroom, lawyers in the courtroom or preoperative surgical explanations, these customizable models resemble real anatomical information. RP from specific subject data allows educational review of the unique clinical variations encountered in pathology.Grant Funding Source: Mentoring Research Grant, College of Life Sciences, Brigham Young University
Signs of persistent neuroinflammation after myocardial infarction of mice (730.3)
Authors: Gouweleeuw Leonie , Pol Christine , Zuidwijk Marianne , Simonides Warner , de Jongste Mike , Eisel Ulrich , Schoemaker Regien 
Published: June 2021

Acute myocardial infarction (MI) evokes production of circulating pro‐inflammatory cytokines, which can result in neuroinflammation. This study aimed to investigate signs of neuroinflammation in mice by measuring brain TNF‐alpha expression and microglia activation after MI.Male 14 week old C57 Bl/6 mice were subjected to MI or sham surgery. Two weeks later, infarct was verified by ECHO‐cardiography, and mice were sacrificed. Brain was processed for western blot on TNF‐alpha, and TNF‐receptors.Heart failure in MI mice was confirmed by increased lung dry weight (35.2±2.2 vs 30.1±0.7 mg), and reduced fractional shortening (19±3 vs 46±1%). TNF‐alpha precursor expression was significantly increased in MI versus sham (9.79±0.60 vs 6.99±0.64 AU) and was associated with higher TNF‐R1 (5.92±2.39 vs 1.88±0.36 AU) and lower TNF‐R2 expression (0.71±0.61 vs 3.01±0.95 AU). Brain tissue stained for microglia showed increased numbers of microglia in the dentate gyrus of the hippocampus (17.1±0.8 vs 14.7±0.8 per high power field) combined with a decreased ratio of processes/cell body surface area (0.071±0.004 vs 0.098±0.007), indicating a higher degree of activation.MI in mice induces increased TNF‐alpha expression at higher TNF‐R1 and lower TNF‐R2 expression in the brain, indicating a persistent pro‐inflammatory state. Spatially altered number of microglia may suggest localization of this inflammation.
A quantitative analysis of skin collagen of Wistar rats submitted to painful procedures in the neonatal period (734.2)
Authors: Carmo Elisabete , Gonçalves Raphaela , Sanada Luciana , Machado Nathália , Violante Izilda , Fazan Valéria Paula 
Published: June 2021

We aimed to quantify skin collagen fibers in animals exposed to painful stimulation after birth. Female Wistar rats were separated into pain group (N=6) and control group (N=6). Pain group animals were stimulated with a needle whereas for controls we used a cotton swab. Stimuli were done on the right paw, twice a day for 15 days since birth. The animals were weighed, anesthetized and killed. The skin of the right and left plantar area of the hindpaws were dissected and prepared for paraffin sections that were Masson trichrome stained. Histological quantification was performed with Image J software. Quantitative data were compared between groups and differences were considered significant when p<0.05. No differences between groups were found for body weight. There was a trend towards a higher collagen percentage on the stimulated paw of the pain group, but no significant difference was found. Also, no difference was found between paws on the control group. Our data suggest that painful stimulation on neonatal period can increase skin collagen fibers at the stimulated region. An increase in the number of specimens studied will confirm this hypothesis.Grant Funding Source: CAPES, FAPESP, CNPq and FAEPA
QnrVC reducing fluoroquinolone susceptibility in epidemic strains of Vibrio cholerae (735.6)
Authors: Kumar Pramod , Yadav Priti , Biswal Sarmistha , Kumar Dhirendra , Sharma Naresh , Bhuyan Soubhagya , Joshi Pallavi , Jain Meenu , Goel Ajay , Kumari Abha , Santosh Baby , Mishra Deepak , Varshney Akhil , Jaiswal Rishi , Singh Lokendra , Yadava Pramod 
Published: June 2021

Treatment of cholera still is a global challenge for health authorities across the world. It is important to use effective antimicrobials for curbing cholera and shortens the duration of illness. As per the WHO recommendations, ciprofloxacin and doxycycline are drugs of choice to be used in treatment of severe cholera. Single doses of ciprofloxacin have been reported to be better than single dose of doxycycline in eradication of cholera bacterium from stool. Earlier we have reported that reduced susceptibility or resistance to fluoroquinolones in Vibrio cholerae O1 is associated with either delay or failure in cholera treatment. V. cholerae harbours two related type II topoisomerases, which alter DNA topology through a DNA cleavage complex. Fluoroquinolones stabilizing the DNA breakage‐reunion complex inhibit DNA replication. Point mutations in gyraseA and ParC responsible for reduced susceptibility to fluoroquinolones are well known. Here, we discuss the QnrVC mediated acquired mechanism of resistance in epidemic strains of V. cholerae.PCR analyses and DNA sequencing revealed presence of qnrVC gene (encoding QnrVC) in epidemic V. cholerae O1 in strains from central and Eastern parts of India. Higher order structural analyses confirmed that QnrVC exhibits right‐handed quadrilateral beta‐helical fold. The shape, size, and charge distribution on QnrVC is reminiscent of a 30‐bp long B‐form DNA. Docking studies revealed that it occupies the entire length of the G segment DNA binding site of type II topoisomerases, thus interfering with fluoroqunolone action. Strains harbouring the qnrVC elements were either resistant or less susceptible to ciprofloxacin as per CLSI, 2010 USA guidelines. Cloning of qnrVC in E. coli transformant mediated 4‐8 folds increment of MIC for ciprofloxacin. The use fluoroquinolones in cholera cases should be closely monitored and development of alternate chemotherapeutic agents targetting transition type II topoisomerases should be given due priority.Grant Funding Source: UGC, DBT and DST, Govt. of India
ATM regulates glioma migration and invasion primarily via MEK‐ERK and not AKT signaling (735.8)
Authors: Allen Jasmine 
Published: June 2021

Glioblastoma multiforme (GBM) is a lethal brain cancer with a life expectancy of only 12‐15 months. Current standard treatment for GBM is surgery and chemoradiation. However, there are many difficulties when it comes to treating this aggressive brain cancer due to inherent radio‐ and chemo‐resistance. Hence, there is a critical need for new therapeutic approaches to make radiation/chemotherapy more effective. Ataxia telangiectasia (A‐T) patients are extremely sensitive to ionizing radiation. The protein mutated in A‐T, ATM (A‐T mutated), controls the cells’ response to radiation and is referred to as the DNA damage response (DDR). ATM is a master protein kinase that targets numerous proteins during the DDR including regulators of cell cycle checkpoints, DNA repair, and apoptosis. In addition, we have shown that without any radiation, ATM might regulate glioma dispersal. Using a highly specific inhibitor of the ATM kinase (ATMi), we recently showed that ATM regulates both ERK and AKT signaling and controls glioma cell migration and invasion in vitro. In order to determine whether MEK‐ERK and/or AKT signaling regulate migration and invasion downstream of ATM, glioma cells were exposed to a MEKi (PD0325901) or AKTi (MK‐2266) in ‘scratch’ and invasion assays. We found that MEKi inhibited both migration and invasion whereas ATKi did not affect migration and only partially (□40%) inhibited invasion. Then, we used a genetic approach and knocked‐down ATM in human glioma cells. In vitro growth and survival studies showed that ATM KD cells were more radiosensitive and grew slower than control cells. Furthermore, we recapitulated the findings with the ATMi, MEKi, and AKTi and showed that ATM KD cells have reduced ability to migrate and invade in vitro suggesting that ATM regulates glioma migration and invasion primarily via MEK‐ERK and less so via AKT signaling. Ongoing studies are determining whether orthotopic ATM KD tumors grow slower and are less invasive than matched control tumors.Grant Funding Source: Supported by NIH
Specialized human DNA polymerases can incorporate ribonucleotides during lesion bypass and DNA repair (736.5)
Authors: Maga Giovanni , Hübscher Ulrich , van Loon Barbara 
Published: June 2021

During DNA replication, replicative DNA polymerases (pols) can incorporate up to 1,000,000 ribonucleotides (rNTPs) into a mammalian genome, which are subsequently removed by the specialized enzyme RNaseH2. The significance and consequences of the presence of rNTPs into the genome are still not well understood. In addition to replication, also DNA repair is a potential source of rNTPs incorporation, but very little is known on the ability of specialized repair DNA pols to incorporate rNTPs. Here we have analyzed the proficiency of rNTPs incorporation opposite normal and damaged bases by DNA pols λ, β, η and ι. Our results established a hierarchy with respect to deoxy‐ vs. ribo‐nucleotide discrimination among these DNA pols and support the notion that, especially in the presence of strongly unbalanced dNTPs/rNTPs pools, as in resting cells, DNA repair could be a major source or rNTPs incorporation.Grant Funding Source: Supported by Italian Cancer Research Association AIRC
Investigation of lysine acetylation and autophagy in the protozoan parasite Toxoplasma gondii (737.3)
Authors: Varberg Joseph , Arrizabalaga Gustavo , Sullivan William 
Published: June 2021

Toxoplasma gondii is an obligate intracellular parasite belonging to the phylum Apicomplexa that chronically infects approximately one‐third of the world’s population. In particular, T. gondii is an opportunistic pathogen that causes severe comorbidities in AIDS patients and immunocompromised individuals. Despite the high disease burden, current drug therapy options are limited and are associated with high incidences of toxicity. Recent studies on the anti‐parasitic ionophore monensin suggest that this drug acts by inducing an autophagic cell death mechanism in the parasite. Interestingly, the T. gondii homologue of the autophagy protein Atg8 was recently identified as being acetylated in a global analysis of T. gondii protein lysine acetylation. As lysine acetylation has been shown to regulate autophagy in other species, we sought to identify the functional significance of the identified Atg8 acetylation mark in T. gondii. Parasites expressing ectopic forms of Atg8 in which the acetylated lysine was mutated to either arginine (non‐acetylateable mutation) or glutamine (an acetyl‐lysine mimic mutation) show altered Atg8 localization. Additionally, expression of the mutant forms of Atg8 led to morphological changes in the parasites reminiscent of those induced during starvation. Current studies are underway to identify if Atg8 mutants have impaired autophagic clearance, or altered ability to interact with other autophagy proteins.Grant Funding Source: Supported by NIH AI077502 (to WJS) and NIH AI 89808‐01 (to GA)