Reproducible Disc Degeneration Scale in a Large Animal Model

IntroductionTo study the efficacy of novel regenerative strategies is necessary to develop new models that do not implement annulus fibrosus (AF) damage. We hypothesize an ideal preclinical model t..

Common germline variants within the CDKN2A/2B region affect risk of pancreatic neuroendocrine tumors

Pancreatic neuroendocrine tumors (PNETs) are heterogeneous neoplasms which represent only 2% of all pancreatic neoplasms by incidence, but 10% by prevalence. Genetic risk factors could have an important role in the disease aetiology, however only a small number of case control studies have been performed yet. To further our knowledge, we genotyped 13 SNPs belonging to the pleiotropic CDKN2A/B gene region in 320 PNET cases and 4436 controls, the largest study on the disease so far. We observed a statistically significant association between the homozygotes for the minor allele of the rs2518719 SNP and an increased risk of developing PNET (ORhom = 2.08, 95% CI 1.05-4.11, p = 0.035). This SNP is in linkage disequilibrium with another polymorphic variant associated with increased risk of several cancer types. In silico analysis suggested that the SNP could alter the sequence recognized by the Neuron-Restrictive Silencer Factor (NRSF), whose deregulation has been associated with the development of several tumors. The mechanistic link between the allele and the disease has not been completely clarified yet but the epidemiologic evidences that link the DNA region to increased cancer risk are convincing. In conclusion, our results suggest rs2518719 as a pleiotropic CDKN2A variant associated with the risk of developing PNETs

Persistent Megalocystic Ovary Following in Vitro Fertilization in a Postpartum Patient with Polycystic Ovarian Syndrome

SummaryObjectiveOvarian hyperstimulation syndrome (OHSS) is more severe when pregnancy occurs, as the developing pregnancy produces human chorionic gonadotropin, which stimulates the ovary's persistent growth. If no pregnancy occurs, the syndrome will typically resolve within 1 week. In a maintained pregnancy, slow resolution of symptoms usually occurs over 1-2 months.Case ReportA 31-year-old woman, gravida 2, para 1, aborta 1, with polycystic ovary syndrome underwent in vitro fertilization (IVF) with clomiphene citrate and follicle-stimulating hormone/gonadotropin releasing hormone-antagonist stimulation. During transvaginal oocyte retrieval, enlarged bilateral ovaries were noted. She had an episode of OHSS after IVF/embryo transfer, for which paracentesis was performed three times. Pregnancy was achieved. Throughout antenatal examinations, bilateral ovaries were enlarged. She delivered a healthy baby by cesarean section at term. However, 1 month after delivery, the bilateral ovary had not shrunk, and levels of tumor markers CA125 and CA199 were 50.84 and 41.34 U/mL, respectively. At laparotomy for suspected malignancy, both adnexae formed “kissing ovaries”, which were multinodulated with yellow serous fluid. Specimens from wedge resection submitted for frozen section showed a benign ovarian cyst. The final pathology report showed bilateral follicle cysts.ConclusionWith the increasing use of gonadotropins in the management of infertility, ovarian enlargement secondary to hyperstimulation is common. Generally, symptoms appear between the 6th and 13th weeks of pregnancy and disappear thereafter. The hyperstimulated ovary often subsides after the first trimester. This case is unusual as the megalocystic ovary persisted after delivery. To the best of our knowledge, we report the first case of enlarged bilateral ovaries persisting 2 months after delivery

Common genetic variants associated with pancreatic adenocarcinoma may also modify risk of pancreatic neuroendocrine neoplasms (vol 39, pg 360, 2018)

Pancreatic neuroendocrine neoplasms (pNEN) account for less than 5% of all pancreatic neoplasms and genetic association studies on susceptibility to the disease are limited. We sought to identify possible overlap of genetic susceptibility loci between pancreatic ductal adenocarcinoma (PDAC) and pNEN; therefore, PDAC susceptibility variants (n=23) from Caucasian genome-wide association studies (GWAS) were genotyped in 369 pNEN cases and 3,277 controls from the PANcreatic Disease ReseArch (PANDoRA) consortium to evaluate the odds associated with pNEN risk, disease onset and tumor characteristics. Main effect analyses showed four PDAC susceptibility variants – rs9854771, rs1561927, rs9543325 and rs10919791 to be associated with pNEN risk. Subsequently, only associations with rs9543325, rs10919791 and rs1561927 were noteworthy with false positive report probability (FPRP) tests. Stratified analyses considering age at onset (50 year threshold), showed rs2736098, rs16986825 and rs9854771 to be associated with risk of developing pNEN at a younger age. Stratified analyses also showed some SNPs to be associated with different degrees of tumor grade, metastatic potential and functionality. Our results identify known GWAS PDAC susceptibility loci, which may also be involved in sporadic pNEN etiology and suggest that some genetic mechanisms governing pathogenesis of these two entities may be similar, with few of these loci being more influential in younger cases or tumor subtypes

Three new pancreatic cancer susceptibility signals identified on chromosomes 1q32.1, 5p15.33 and 8q24.21.

Genome-wide association studies (GWAS) have identified common pancreatic cancer susceptibility variants at 13 chromosomal loci in individuals of European descent. To identify new susceptibility variants, we performed imputation based on 1000 Genomes (1000G) Project data and association analysis using 5,107 case and 8,845 control subjects from 27 cohort and case-control studies that participated in the PanScan I-III GWAS. This analysis, in combination with a two-staged replication in an additional 6,076 case and 7,555 control subjects from the PANcreatic Disease ReseArch (PANDoRA) and Pancreatic Cancer Case-Control (PanC4) Consortia uncovered 3 new pancreatic cancer risk signals marked by single nucleotide polymorphisms (SNPs) rs2816938 at chromosome 1q32.1 (per allele odds ratio (OR) = 1.20, P = 4.88x10 -15), rs10094872 at 8q24.21 (OR = 1.15, P = 3.22x10 -9) and rs35226131 at 5p15.33 (OR = 0.71, P = 1.70x10 -8). These SNPs represent independent risk variants at previously identified pancreatic cancer risk loci on chr1q32.1 ( NR5A2), chr8q24.21 ( MYC) and chr5p15.33 ( CLPTM1L- TERT) as per analyses conditioned on previously reported susceptibility variants. We assessed expression of candidate genes at the three risk loci in histologically normal ( n = 10) and tumor ( n = 8) derived pancreatic tissue samples and observed a marked reduction of NR5A2 expression (chr1q32.1) in the tumors (fold change -7.6, P = 5.7x10 -8). This finding was validated in a second set of paired ( n = 20) histologically normal and tumor derived pancreatic tissue samples (average fold change for three NR5A2 isoforms -31.3 to -95.7, P = 7.5x10 -4-2.0x10 -3). Our study has identified new susceptibility variants independently conferring pancreatic cancer risk that merit functional follow-up to identify target genes and explain the underlying biology

Genetic determinants of telomere length and risk of pancreatic cancer: A PANDoRA study

Telomere deregulation is a hallmark of cancer. Telomere length measured in lymphocytes (LTL) has been shown to be a risk marker for several cancers. For pancreatic ductal adenocarcinoma (PDAC) consensus is lacking whether risk is associated with long or short telomeres. Mendelian randomization approaches have shown that a score built from SNPs associated with LTL could be used as a robust risk marker. We explored this approach in a large scale study within the PANcreatic Disease ReseArch (PANDoRA) consortium. We analyzed 10 SNPs (ZNF676-rs409627, TERT-rs2736100, CTC1-rs3027234, DHX35-rs6028466, PXK-rs6772228, NAF1-rs7675998, ZNF208-rs8105767, OBFC1-rs9420907, ACYP2-rs11125529 and TERC-rs10936599) alone and combined in a LTL genetic score (“teloscore”, which explains 2.2% of the telomere variability) in relation to PDAC risk in 2,374 cases and 4,326 controls. We identified several associations with PDAC risk, among which the strongest were with the TERT-rs2736100 SNP (OR = 1.54; 95%CI 1.35–1.76; p = 1.54 × 10−10) and a novel one with the NAF1-rs7675998 SNP (OR = 0.80; 95%CI 0.73–0.88; p = 1.87 × 10−6, ptrend = 3.27 × 10−7). The association of short LTL, measured by the teloscore, with PDAC risk reached genome-wide significance (p = 2.98 × 10−9 for highest vs. lowest quintile; p = 1.82 × 10−10 as a continuous variable). In conclusion, we present a novel genome-wide candidate SNP for PDAC risk (TERT-rs2736100), a completely new signal (NAF1-rs7675998) approaching genome-wide significance and we report a strong association between the teloscore and risk of pancreatic cancer, suggesting that telomeres are a potential risk factor for pancreatic cancer

IN VIVO CHARACTERIZATION OF A NEW TRANSPEDICULAR APPROACH TO THE INTERVERTEBRAL DISC IN A LARGE ANIMAL MODEL

Back pain (BP) is a common clinical condition that leads to high morbidity with significant psychosocial and economic effects. It is the leading cause of disability in people under 45 years of age, and it results in enormous national economic losses in developed countries. The wide majority of BP is associated with degenerative changes of the intervertebral disc (IVD). Intervertebral disc degeneration (IVDD) is an age-related chronic process that is characterized by a progressive reduction of proteoglycans and water content in the nucleus pulposus (NP) with loss of the IVD's ability to resist compressive forces. Current treatment options for BP and IVDD range from conservative measures to invasive procedures however, these treatment modalities have limited efficacy and do not produce predictable and reliable outcomes. Therefore, there is a clear need for more effective early treatments of BP that may prevent, slow down, or reverse the degenerative changes. To evaluate the efficacy of novel IVD regenerative treatments, prior to translation into humans, ex vivo and in vivo animal model systems are needed. In the present study, a novel transpedicular approach to the IVD has been validated in 12 sheep. Under fluoroscopy, a 2-mm Kirshner wire was introduced through the vertebral body with a inclination of approximately 45° in all plans direction to reach the center of the NP. In each animal, four IVDs, from L1 to L5, were addressed by performing different surgical techniques respectively: (I) nucleotomy, (II) tunnel, (III) nucleotomy + polyurethane (PU) scaffold, and (IV) tunnel + PU scaffold. Intact IVDs (L5-6) were used as controls. Intra- and post-surgical morbidity rates were low; CSF leakage was recorded in two spinal segments whereas discospondylitis and vertebral luxation occurred respectively at L1-2 and L3-4 in two different animals. The quantitative and qualitative analysis of MRI, radiologic, histologic and macroscopic data, collected at four different time points (before and 1, 3 and 6 months after surgery), suggested that the injury induced in the present model represents a reliable method for initiating a progressive IVDD process, obtaining different degrees of IVDD depending on the type of lesion performed. The endplate damage itself, caused by the realization of the transpedicular tunnel, led to IVD degenerative changes, although to a lesser extent than those caused by performing nucleotomy. Furthermore, the sealing of the tunnel with the PU scaffold resulted in a lack of cells leakage throughout the tunnel. The transpedicular approach represents a feasible alternative route to the IVD, without causing damage to the annulus fibrosus. This new pathway to the IVD provides a new valid model to study biologic and biomechanical alterations in relation to both IVD degenerative processes and potential NP regenerative therapies.La lombalgia è una condizione clinica molto frequente associata ad elevata morbidità, con effetti sia dal punto di vista psicosociale che della spesa sanitaria nazionale. La lombalgia é la principale causa di disabilità nella popolazione al di sotto dei 45 anni e rappresenta una della principali voci di spesa nei paesi sviluppati. La grande maggioranza dei casi di lombalgia è associata alla degenerazione del disco intervertebrale (IVDD). L'IVDD è un processo cronico età-correlato caratterizzato da una progressiva riduzione del contenuto di proteoglicani ed acqua nel nucleo polposo (NP), con perdita della capacità del disco di resistere a forze compressive. Le attuali opzioni di trattamento disponibili per lombalgia e IVDD comprendono sia approcci conservativi che procedure invasive. Tuttavia, queste modalità di trattamento hanno un' efficacia limitata e non producono risultati prevedibili e riproducibili. Per queste ragioni, vi è una evidente necessità di trattamenti che siano efficaci nel prevenire, rallentare o invertire le modificazioni degenerative causate dal processo di IVDD. Al fine di valutare l’efficacia di nuove tecniche di rigenerazione discale e prima di trasferirle all’uomo, é necessario testarle su modelli animali ex vivo ed in vivo. In questo studio, un nuovo approccio al disco intervertebrale (IVD) per via transpeduncolare, é stato valicato in 12 pecore. Tramite guida fluoroscopica, un filo di Kirshner di 2-mm di diametro é stato introdotto attraverso il corpo vertebrale con un’ inclinazione di circa 45° in tutti i piani dello spazio, tale da raggiungere il centro del NP. In ciascun animale sono stati trattati quattro IVD, da L1 a L5, effettuando in ciascuno una diversa tecnica chirurgica: (I) nucleotomia, (II) tunnel, (III) nucleotomia + scaffold in poliuretano (PU) e (IV) tunnel + scaffold PU. I dischi non trattati (L5-6) sono stati considerati come gruppo controllo. Le percentuali di morbiditá intra- e post-operatoria si sono rivelate basse; in due occasioni é stata riportata fuoriuscita di liquor in fase chirurgica, inoltre lussazione di L3-4 e discospondilite a livello di L1-2 si sono verificate in due soggetti. Le analisi quantitative e qualitative, effettuate sui dati ricavati in quattro diversi time point (prima e 1, 3 e 6 mesi dopo la chirurgia) dalle immagini radiografiche, di risonanza magnetica e dai campioni macroscopici e istologici, hanno rivelato l’efficacia del metodo presentato nell’ottenere un modello progressive di IVDD. Attraverso l’approccio transpeduncolare é stato possibile ottenere diversi gradi di IVDD, dipendentemente dalla tipologia di lesione effettuata. Il solo danno al piatto vertebrale ha determinato alterazioni degenerative del IVD, tuttavia inferiori rispetto a quelle causate effettuando la nucleotomia. Inoltre, l’utilizzo dello scaffold in PU per sigillare il tunnel é servito a prevenire la fuoriuscita di cellule attraverso il tunnel. L’approccio transpeduncolare rappresenta una via alternativa per raggiungere il IVD, senza danneggiare l’anello fibroso. Questa nuova tecnica fornisce un nuovo modello per lo studio di alterazioni biologiche e biomeccaniche in relazione sia ai processi di IVDD sia a potenziali terapie regenerative

IN VIVO CHARACTERIZATION OF A NEW TRANSPEDICULAR APPROACH TO THE INTERVERTEBRAL DISC IN A LARGE ANIMAL MODEL

Back pain (BP) is a common clinical condition that leads to high morbidity with significant psychosocial and economic effects. It is the leading cause of disability in people under 45 years of age, and it results in enormous national economic losses in developed countries. The wide majority of BP is associated with degenerative changes of the intervertebral disc (IVD). Intervertebral disc degeneration (IVDD) is an age-related chronic process that is characterized by a progressive reduction of proteoglycans and water content in the nucleus pulposus (NP) with loss of the IVD's ability to resist compressive forces. Current treatment options for BP and IVDD range from conservative measures to invasive procedures however, these treatment modalities have limited efficacy and do not produce predictable and reliable outcomes. Therefore, there is a clear need for more effective early treatments of BP that may prevent, slow down, or reverse the degenerative changes. To evaluate the efficacy of novel IVD regenerative treatments, prior to translation into humans, ex vivo and in vivo animal model systems are needed. In the present study, a novel transpedicular approach to the IVD has been validated in 12 sheep. Under fluoroscopy, a 2-mm Kirshner wire was introduced through the vertebral body with a inclination of approximately 45° in all plans direction to reach the center of the NP. In each animal, four IVDs, from L1 to L5, were addressed by performing different surgical techniques respectively: (I) nucleotomy, (II) tunnel, (III) nucleotomy + polyurethane (PU) scaffold, and (IV) tunnel + PU scaffold. Intact IVDs (L5-6) were used as controls. Intra- and post-surgical morbidity rates were low; CSF leakage was recorded in two spinal segments whereas discospondylitis and vertebral luxation occurred respectively at L1-2 and L3-4 in two different animals. The quantitative and qualitative analysis of MRI, radiologic, histologic and macroscopic data, collected at four different time points (before and 1, 3 and 6 months after surgery), suggested that the injury induced in the present model represents a reliable method for initiating a progressive IVDD process, obtaining different degrees of IVDD depending on the type of lesion performed. The endplate damage itself, caused by the realization of the transpedicular tunnel, led to IVD degenerative changes, although to a lesser extent than those caused by performing nucleotomy. Furthermore, the sealing of the tunnel with the PU scaffold resulted in a lack of cells leakage throughout the tunnel. The transpedicular approach represents a feasible alternative route to the IVD, without causing damage to the annulus fibrosus. This new pathway to the IVD provides a new valid model to study biologic and biomechanical alterations in relation to both IVD degenerative processes and potential NP regenerative therapies.La lombalgia è una condizione clinica molto frequente associata ad elevata morbidità, con effetti sia dal punto di vista psicosociale che della spesa sanitaria nazionale. La lombalgia é la principale causa di disabilità nella popolazione al di sotto dei 45 anni e rappresenta una della principali voci di spesa nei paesi sviluppati. La grande maggioranza dei casi di lombalgia è associata alla degenerazione del disco intervertebrale (IVDD). L'IVDD è un processo cronico età-correlato caratterizzato da una progressiva riduzione del contenuto di proteoglicani ed acqua nel nucleo polposo (NP), con perdita della capacità del disco di resistere a forze compressive. Le attuali opzioni di trattamento disponibili per lombalgia e IVDD comprendono sia approcci conservativi che procedure invasive. Tuttavia, queste modalità di trattamento hanno un' efficacia limitata e non producono risultati prevedibili e riproducibili. Per queste ragioni, vi è una evidente necessità di trattamenti che siano efficaci nel prevenire, rallentare o invertire le modificazioni degenerative causate dal processo di IVDD. Al fine di valutare l’efficacia di nuove tecniche di rigenerazione discale e prima di trasferirle all’uomo, é necessario testarle su modelli animali ex vivo ed in vivo. In questo studio, un nuovo approccio al disco intervertebrale (IVD) per via transpeduncolare, é stato valicato in 12 pecore. Tramite guida fluoroscopica, un filo di Kirshner di 2-mm di diametro é stato introdotto attraverso il corpo vertebrale con un’ inclinazione di circa 45° in tutti i piani dello spazio, tale da raggiungere il centro del NP. In ciascun animale sono stati trattati quattro IVD, da L1 a L5, effettuando in ciascuno una diversa tecnica chirurgica: (I) nucleotomia, (II) tunnel, (III) nucleotomia + scaffold in poliuretano (PU) e (IV) tunnel + scaffold PU. I dischi non trattati (L5-6) sono stati considerati come gruppo controllo. Le percentuali di morbiditá intra- e post-operatoria si sono rivelate basse; in due occasioni é stata riportata fuoriuscita di liquor in fase chirurgica, inoltre lussazione di L3-4 e discospondilite a livello di L1-2 si sono verificate in due soggetti. Le analisi quantitative e qualitative, effettuate sui dati ricavati in quattro diversi time point (prima e 1, 3 e 6 mesi dopo la chirurgia) dalle immagini radiografiche, di risonanza magnetica e dai campioni macroscopici e istologici, hanno rivelato l’efficacia del metodo presentato nell’ottenere un modello progressive di IVDD. Attraverso l’approccio transpeduncolare é stato possibile ottenere diversi gradi di IVDD, dipendentemente dalla tipologia di lesione effettuata. Il solo danno al piatto vertebrale ha determinato alterazioni degenerative del IVD, tuttavia inferiori rispetto a quelle causate effettuando la nucleotomia. Inoltre, l’utilizzo dello scaffold in PU per sigillare il tunnel é servito a prevenire la fuoriuscita di cellule attraverso il tunnel. L’approccio transpeduncolare rappresenta una via alternativa per raggiungere il IVD, senza danneggiare l’anello fibroso. Questa nuova tecnica fornisce un nuovo modello per lo studio di alterazioni biologiche e biomeccaniche in relazione sia ai processi di IVDD sia a potenziali terapie regenerative

Lumbosacral intraspinal extradural ganglion cyst in a cat

Case summary A 16-year-old neutered female domestic shorthair cat was referred for chronic history of reluctance to jump, stiffness of the tail and lower back pain. Mild pelvic limb ataxia, reduced perianal reflex and lumbosacral discomfort were present on neurological examination. On magnetic resonance imaging, a well-defined rounded structure of 3 mm in diameter was identified on the right dorsal aspect of the epidural space at L7–S1, causing displacement of the cauda equina. The lesion was hyperintense to spinal cord parenchyma on T2-weighted images and hypointense on T1-weighted images, consistent with a fluid-filled structure. A Lumbosacral dorsal laminectomy was performed. A clear fluid-containing structure was identified between the right L7 nerve root and the cauda equina. Following surgical excision, histopathology confirmed the cystic nature of the lesion and revealed thick disorganised sheaths of fibrocollagenous tissue and flattened mesenchymal cells lining the luminal part of the cyst wall. A diagnosis of intraspinal ganglion cyst was made. The cat recovered uneventfully. Seven months after surgery euthanasia was performed for unrelated reasons; no neurological deficits were present. Relevance and novel information This is the first reported case of intraspinal ganglion cyst in a cat. Intraspinal extradural cysts should be considered among other differential diagnoses for cats with lumbosacral myelopathy/radiculopathy