Tail Necrosis Research Articles

S1937 Acute Pancreatitis Due to Vascular Insufficiency From Celiac Artery Stenosis

Introduction: Acute pancreatitis is a leading cause of hospitalizations in the United States. The pancreas relies on blood supply from the celiac and superior mesenteric arteries. Compromise to any part of the pancreatic circulation may lead to inflammation. Vascular insufficiencies are an uncommon cause of acute pancreatitis. We present a rare case of acute pancreatitis secondary to celiac stenosis with concomitant abdominal aortic aneurysm thrombosis. Case Description/Methods: The patient is a 79-year-old female with history of tobacco use and peripheral vascular disease that presented for one day of abdominal pain. She noted nausea with non-bloody emesis during this time. Labs revealed a leukocytosis with WBC 17.1, lactate 2.7, AST 25, ALT 16, alkaline phosphatase 53, Total bilirubin 0.8, direct bilirubin 0.1, and a lipase of 976. A CTA of the abdomen was demonstrated inflammatory fluid with pancreatic tail necrosis, peripancreatic stranding, stenosis of the celiac origin and thrombosed aortic aneurysm measuring 6.1cm with distal reconstitution. She was placed on intravenous heparin and lactated ringer infusions. After the diagnosis of her acute pancreatitis, she was found to have a normal triglyceride level and IgG4 level. An abdominal ultrasound revealed gallbladder sludge without cholelithiasis or biliary dilation. There was no history of alcohol use preceding her hospitalization. Moreover, there were no new medications or over the counter supplements reported. It was felt the patient’s pancreatitis was a result of arterial insufficiency from celiac artery stenosis. The patient was seen by vascular surgery and was treated medically. During her stay she developed worsening pleural effusion and hypoxic respiratory failure requiring oxygen. She was ultimately discharged to a skilled nursing facility when medically stable (Figure). Discussion: Isolated case reports of acute pancreatitis due to vascular insufficiencies have been reported without defined incidence or prevalence. The head of the pancreas receives blood supply from the celiac trunk and superior mesenteric artery respectively. The body and tail of the pancreas are more vulnerable to ischemia, being perfused solely by the splenic artery. After ruling out common etiologies, compromise of the celiac artery was presumed to be the cause of this patient’s disease process. Conservative treatment yielded satisfactory results in our case, however the standard of care regarding acute pancreatitis with vascular insufficiencies need additional elucidation.Figure 1.: A. Celiac artery stenosis. B. Pancreatitis with pancreatic tail necrosis and peripancreatic fluid and stranding. C. Thrombosis within distal aorta.

Curcuminoid Co-Loading Platinum Heparin-Poloxamer P403 Nanogel Increasing Effectiveness in Antitumor Activity.

Nanosized multi-drug delivery systems provide synergistic effects between drugs and bioactive compounds, resulting in increased overall efficiency and restricted side effects compared to conventional single-drug chemotherapy. In this study, we develop an amphiphilic heparin-poloxamer P403 (HP403) nanogel that could effectively co-load curcuminoid (Cur) and cisplatin hydrate (CisOH) (HP403@CisOH@Cur) via two loading mechanisms. The HP403 nanogels and HP403@CisOH@Cur nanogels were closely analyzed with 1H-NMR spectroscopy, FT-IR spectroscopy, TEM, and DLS, exhibiting high stability in spherical forms. In drug release profiles, accelerated behavior of Cur and CisOH at pH 5.5 compared with neutral pH was observed, suggesting effective delivery of the compounds in tumor sites. In vitro studies showed high antitumor activity of HP403@CisOH@Cur nanogels, while in vivo assays showed that the dual-drug platform prolonged the survival time of mice and prevented tail necrosis. In summary, HP403@CisOH@Cur offers an intriguing strategy to achieve the cisplatin and curcumin synergistic effect in a well-designed delivery platform that increases antitumor effectiveness and overcomes undesired consequences caused by cisplatin in breast cancer treatment.

Sodium chloride injection to treat opioid overdose; Does it work? A preclinical study

Opioid overdoses (ODs) are increasing in Mexico's northern border. Because naloxone is usually not available, witnesses inject common salt (NaCl) into a vein of OD victims in an attempt to help them regain consciousness. Despite this widespread practice, no preclinical studies have addressed the efficacy of NaCl as an opioid antidote. Here we tested saline solutions at different concentrations. Because the highest (31.6 %) caused tail necrosis, we selected 17.7 % as a hypertonic saline solution (HSS) to determine if it could prevent the lethal effect of morphine (Mor), fentanyl (Fen), or Mor + Fen in adult Wistar male rats. We also evaluated if NaCl could modify the opioid antagonist effect of naloxone. Our results show that HSS: a) sensitizes animals to thermal but not mechanical stimuli; b) does not prevent mortality caused by high morphine or fentanyl doses; c) decreases the latency to recovery from the sedative effects caused by low doses of morphine or fentanyl; and d) increases naloxone's efficacy to prevent the lethality produced by Mor or Fen, but not by Mor + Fen. These results suggest that HSS is marginally effective in shortening the recovery time from nonfatal opioid ODs and increases naloxone's efficacy to counteract opioid-induced ODs.

A Murine Tail Lymphedema Model.

Lymphedema is extremity swelling caused by lymphatic dysfunction. The affected limb enlarges because of accumulation of fluid, adipose, and fibrosis. There is no cure for this disease. A mouse tail model that uses a focal full thickness skin excision near the base of the tail, resulting in tail swelling, has been used to study lymphedema. However, this model may result in vascular comprise and consequent tail necrosis and early tail swelling resolution, limiting its clinical translatability. The chronic murine tail lymphedema model induces sustained lymphedema over 15 weeks and a reliable perfusion to the tail. Enhancements of the traditional murine tail lymphedema model include 1) precise full thickness excision and lymphatic clipping using a surgical microscope, 2) confirmation of post-operative arterial and venous perfusion using high resolution laser speckle, and 3) functional assessment using indocyanine green near infrared laser lymphangiography. We also use tissue nanotransfection technology (TNT) for novel non-viral, transcutaneous, focal delivery of genetic cargo to the mouse tail vasculature.

Vitamin D Toxicosis in a Blue-Tongued Skink (Tiliqua scincoides) Presented with Epistaxis and Tongue Discoloration

A 10-yr-old intact blue-tongued skink (Tiliqua scincoides) of unknown sex presented for epistaxis and dyspnea. On physical examination, dysecdysis, necrosis of the tip of the tail, and a pink discoloration of the proximal third of the tongue were noted in addition to epistaxis, dyspnea, and tachypnea. Clinical pathology findings included marked polychromatophilia, moderate heterophilia with signs of toxicity, a regenerative left shift, and moderate hyperglycemia. Because of the suspicion of an infectious etiology, antibiotics were prescribed. Meloxicam was also administered without clinical improvement. Swelling of the pharyngeal area was noted on whole-body computed tomographic and magnetic resonance imaging. Oro-pharyngeal endoscopic-guided biopsies were performed under general anesthesia. Histology was consistent with moderate erosion and goblet cell hyperplasia. Plasma 25-hydroxyvitamin D3 concentration was 768 nmol/L and was interpreted as possible vitamin D toxicosis. Prednisolone was prescribed to enhance calciuresis and treat potential vasculitis, but the skink was ultimately euthanized. Histopathologic examination was consistent with vitamin D toxicosis. The diet of this skink mostly consisted of greens dusted with a supplement containing calcium and vitamin D3. This case report describes the challenges associated with antemortem diagnosis of a vitamin D toxicosis, even with current imaging techniques. It highlights that specific reference intervals should be established for 25-hydroxyvitamin D3 and ionized calcium concentrations in various reptile species. Clinicians presented with a blue-tongued skink displaying a pink tongue or tail necrosis should consider vitamin D toxicosis in their differential diagnosis.

Abstract No. 438 Tail artery access for transarterial experiments in rats: feasibility study in a survival model

The rat is an increasingly valuable model to study interventional therapeutic procedures. Suitable arterial access is essential for vascular intervention. In rats, transtail artery access has been recently reported as a feasible technique compared to conventional carotid or femoral access. The aim of this study was to evaluate repeat accessibility and complications of transtail angiography in a survival model. Seventeen male Sprague-Dawley rats were used (460.2 ± 45.0g), one rat died before the procedure, hence 16 rats were included in this study. 1-cm longitudinal ventral incisions were created 5 cm distal to the base of the tail to dissect the tail artery (ventral caudal artery). A 1.5–1.7 Fr microcatheter was advanced over a 0.010–0.014 inch microguidewire through a partial arteriotomy. Selective renal angiography was then performed. Intraarterial antibiotics injection was performed in 11 of 16 rats. The accessed segment of the tail artery was ligated and the incision closed with sutures. Repeat renal angiography via the tail artery was performed 48 hours after the initial procedure using a separate tail incision 1 cm proximal to the initial incision. Procedure time from skin incision to catheterization was recorded. Complications including tail ischemia, infection and bleeding were evaluated during the 2-day follow-up period. All initial and repeat procedures were successfully performed. The average time for the 32 tail artery cannulations was 11.0 ± 4.5 min. Transient focal skin color change in the tail was observed in one rat immediately after the initial procedure, which resolved within 90 mins. No tail necrosis, major bleeding or signs of infection were observed. Dehiscence of the incision was observed in 3 of 16 rats from self-grooming, with no bleeding or infection. Repeat tail artery access for angiography in rats was successful without major complications in a survival model. This reproducible technique can provide a new experimental platform to test transarterial procedures.

Development and Non-clinical Evaluation of an 111In/225Ac Theranostic for Triple Negative Breast Cancer

The purpose of this study was to develop an Indium-111/Actinium-225 based theragnostic utilizing the tMUC1 targeting humanized antibody, TAB004, and its evaluation in a tumor bearing rodent model. Triple negative breast cancer (TNBC) is an aggressive phenotype that comprises only 15-20% of all new breast cancer cases but is responsible for a majority of related deaths. Chemotherapy is the current mainstay treatment for TNBC but side effects and drugresistance remain an issue, so development of targeted therapies for TNBC is critical. Tumor associated Mucin 1 (tMUC1), an aberrantly glycosylated transmembrane glycoprotein, is overexpressed in >90% of TNBCs and as such is an attractive target. The humanized antibody hTAB004 shows high affinity and selectivity to tMUC1 and was thus used to develop a theranostic strategy for TNBC. hTAB004 was conjugated to DOTA-NHS, purified, formulated in a metal free buffer and radiolabeled with Indium-111 or Actinium-225 to produce 111In-DOTA-hTAB004 and 225Ac-DOTA-hTAB004, respectively. The radiolabeled molecule was evaluated for stability. Retention of affinity of cold-labeled molecules was validated. In vivo and ex vivo biodistribution studies of 111In-DOTA-hTAB004 were performed in HCC70 orthotopic xenograft tumor-bearing female NSG mice (n=3) using SPECT/CT imaging over 120h. Dosimetry analysis was performed utilizing the in vivo data. Therapeutic efficacy of 225Ac-DOTA-hTAB004 was determined in HCC70 orthotopic xenograft tumorbearing female nude mice (n=5/group) by monitoring bodyweights and tumor sizes over time. DOTA-hTAB004 was labeled successfully with both Indium-111 and Actinium-225. The radiolabeled molecules were found stable in both formulation and mouse serum and the non-radioactively labeled surrogates 115In-DOTA-hTAB004 and 139La-DOTA-hTAB004 retained their affinity to tMUC1.In vivo biodistribution data revealed increased tumor accumulation of 111In-DOTA-hTAB004 over 120 h, reaching 65±15 percent of injected dose per gram (%ID/g). The next organs of significant uptake (Spleen 8.9±0.6 %ID/g and Liver 8.3±1 %ID/g) had a 7-fold lower uptake at 120 h. All mice treated with a single injection of 225Ac-DOTA-hTAB004 (500 nCi, 18 kBq) showed a complete response with tumor volumes shrinking by > 89% within 48 days. No therapy associated toxicities were seen in any of the mice outside of tail necrosis in one animal which may have been due to dose extravasation. All the control mice, which received the DOTA-hTAB004, showed continued tumor growth and had to be sacrificed by day 35. 111In-DOTA-hTAB004 biodistribution data indicates the excellent tumor targeting capabilities of hTAB004. 225Ac-DOTA-hTAB004 therapy data shows exceptional clinical benefit in mice bearing human tumors. Given this data, radiolabeled hTAB004 is a promising targeted treatment and imaging option for TNBC.

3033 Acute Pancreatitis Complicated by Pancreatic Pseudocyst and Splenic Hematoma, Management Considerations

INTRODUCTION: Pancreatic duct disruption is a known complication of pancreatitis which leads to the formation of pancreatic pseudocysts, pancreatic ascites, and high amylase pleural effusions. Pancreatic pseudocysts migrate along with the pancreatic tissue and rupture either into surrounding organs or into the free peritoneal cavity. Splenic complications of pancreatitis are rare and include splenic vein thrombosis, splenic artery rupture, splenic rupture or infarction, and splenic hematoma. CASE DESCRIPTION/METHODS: 39 y/o alcoholic male admitted for acute pancreatitis. Imaging revealed pseudocysts and ascites. Initially, he improved with supportive care but abdominal pain recurred owing to splenic rupture with resultant splenic hematoma. Splenic artery coiling was employed to stop bleeding. He returned for recurrent abdominal pain and new chest pain with interval increase in size of pseudocyst and interval development of pleural effusion. EUS revealed a multiloculated pancreatic pseudocyst, as well as pancreatic duct disruption. Transgastric drainage of the pseudocyst was performed with an Axios stent. Repeat endoscopy at interval showed complete resolution of the cyst. DISCUSSION: Studies estimate the incidence of pancreatic pseudocyst rupture into the spleen at a rate of up to 1.1% of patients with acute pancreatitis. Another study of patients with chronic pancreatitis reports splenic pseudocyst as the most common lesion, followed by splenic rupture and hematoma. A more recent study of 500 men with chronic alcoholic pancreatitis found that only 2.2% develop splenic complications with intrasplenic pseudocyst being the most common lesion, followed by splenic rupture and subcapsular hematoma. Findings associated with the highest risk of developing splenic complications included pancreatic tail necrosis, distal pseudocyst, or splenic vein occlusion. Previously, the mainstay of treatment for splenic hematoma was splenectomy with distal pancreatectomy. However, with the advent of advanced endoscopic procedures, patients are safely managed with splenic artery embolizationand EUS guided pancreaticogastrostomy or percutaneous drainage. Endoscopic drainage and percutaneous drainage of symptomatic pseudocysts are preferred to surgical management. Furthermore, recent reviews show that endoscopic drainage is superior to percutaneous drainage with lower rates of reintervention, shorter hospital stays, and need for less follow-up imaging.

Abstract LB-281: Development and non-clinical evaluation of an111In/225Ac theranostic for triple negative breast cancer

Abstract Purpose: The purpose of this study was to develop an Indium-111/Actinium-225 based theragnostic utilizing the tMUC1 targeting humanized antibody, TAB004, and its evaluation in a tumor bearing rodent model. Triple negative breast cancer (TNBC) is an aggressive phenotype that comprises only 15-20% of all new breast cancer cases but is responsible for a majority of related deaths. Chemotherapy is the current mainstay treatment for TNBC but side effects and drug-resistance remain an issue, so development of targeted therapies for TNBC is critical. Tumor associated Mucin 1 (tMUC1), an aberrantly glycosylated transmembrane glycoprotein, is overexpressed in >90% of TNBCs and as such is an attractive target. The humanized antibody hTAB004 shows high affinity and selectivity to tMUC1 and was thus used to develop a theranostic strategy for TNBC. Method: hTAB004 was conjugated to DOTA-NHS, purified, formulated in a metal free buffer and radiolabeled with Indium-111 or Actinium-225 to produce 111In-DOTA-hTAB004 and 225Ac-DOTA-hTAB004, respectively. The radiolabeled molecule was evaluated for stability. Retention of affinity of cold-labeled molecules was validated. In vivo and ex vivo biodistribution studies of 111In-DOTA-hTAB004 were performed in HCC70 orthotopic xenograft tumor-bearing female NSG mice (n=3) using SPECT/CT imaging over 120h. Dosimetry analysis was performed utilizing the in vivo data. Therapeutic efficacy of 225Ac-DOTA-hTAB004 was determined in HCC70 orthotopic xenograft tumor-bearing female nude mice (n=5/group) by monitoring bodyweights and tumor sizes over time. Results: DOTA-hTAB004 was labeled successfully with both Indium-111 and Actinium-225. The radiolabeled molecules were found stable in both formulation and mouse serum and the non-radioactively labeled surrogates 115In-DOTA-hTAB004 and 139La-DOTA-hTAB004 retained their affinity to tMUC1. In vivo biodistribution data revealed increased tumor accumulation of 111In-DOTA-hTAB004 over 120 h, reaching 65±15 percent of injected dose per gram (%ID/g). The next organs of significant uptake (Spleen 8.9±0.6 %ID/g and Liver 8.3±1 %ID/g) had a 7-fold lower uptake at 120 h. All mice treated with a single injection of 225Ac-DOTA-hTAB004 (500 nCi, 18 kBq) showed a complete response with tumor volumes shrinking by > 89% within 48 days. No therapy associated toxicities were seen in any of the mice outside of tail necrosis in one animal which may have been due to dose extravasation. All the control mice, which received the DOTA-hTAB004, showed continued tumor growth and had to be sacrificed by day 35. Conclusions 111In-DOTA-hTAB004 biodistribution data indicates the excellent tumor targeting capabilities of hTAB004. 225Ac-DOTA-hTAB004 therapy data shows exceptional clinical benefit in mice bearing human tumors. Given this data, radiolabeled hTAB004 is a promising targeted treatment and imaging option for TNBC. Citation Format: ohad ilovich, Vanessa Kelly, Shu-ta Wu, Vijay Gottumukkala, Richard Coelho, Rahul Puri, Pinku Mukherjee. Development and non-clinical evaluation of an111In/225Ac theranostic for triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr LB-281.

Expression of a constitutively active human STING mutant in hematopoietic cells produces an Ifnar1-dependent vasculopathy in mice.

STING-associated vasculopathy with onset in infancy (SAVI) is an autoinflammatory disorder characterized by blood vessel occlusions, acral necrosis, myositis, rashes, and pulmonary inflammation that are the result of activating mutations in the STimulator of Interferon Genes (STING). We generated a transgenic line that recapitulates many of the phenotypic aspects of SAVI by targeting the expression of the human STING-N154S-mutant protein to the murine hematopoietic compartment. hSTING-N154S mice demonstrated failure to gain weight, lymphopenia, progressive paw swelling accompanied by inflammatory infiltrates, severe myositis, and ear and tail necrosis. However, no significant lung inflammation was observed. X-ray microscopy imaging revealed vasculopathy characterized by arteriole occlusions and venous thromboses. Type I interferons and proinflammatory mediators were elevated in hSTING-N154S sera. Importantly, the phenotype was prevented in hSTING-N154S mice lacking the type I interferon receptor gene (Ifnar1). This model, based on a mutant human STING protein, may shed light on the pathophysiological mechanisms operative in SAVI.

A modified protocol for generating the simulated weightlessness rat model

To introduce a modified protocol for generating the simulated weightlessness rat model by hindlimb unloading. Ninety male adult SD rats were randomly divided into three groups: the control group, classical suspension group and modified suspension group (n=30/group). In the classical suspension group, a strip of medical adhesive tape was attached to the tail, with horizontal filament tape wrapping. A piece of gauze was wrapped around the tail at the outermost layer and the tail was suspended for hindlimb unloading. In the modified suspension group, a layer of plastic net was added between the horizontal filament tape and the gauze to reduce the squeeze on the tail as a buffer zone and ensure proper circulation of the tail. After 4 weeks of suspension, damage to the tail and sheath detachment were observed. Meanwhile the body weight and right soleus wet weight of rats were measured. The ratio of right soleus wet weight to body weight was decreased significantly in both the classical suspension group and the modified suspension group compared with the control group, while there was no difference in body weight among the three different groups. Importantly, the incidence of tail ischemia and necrosis (13.3% vs 40.0% in the classical suspension group) and the incidence of sheath detachment from tail (3.3% vs 26.7% in the classical suspension group) were significantly lower whereas the success rates of model (33.3% vs 83.3% in classical suspension group) was significantly higher in the modified suspension group. The modified protocol decreases the incidence of tail necrosis and sheath detachment in the rat tail suspension and increases the success rate of the hindlimb unloading rat model, with improved simplicity and practicability.

Tail lesions in fattening pigs: relationships with postmortem meat inspection and influence of a tail biting management tool

Tail biting is a major welfare and economic problem in intensive pig production. The objectives of this study were to (1) determine tail lesion prevalence at a German abattoir, (2) test for associations between meat inspection findings and tail lesions, (3) assess the agreement between tail necrosis recorded during meat inspection and scored from pictures and (4) test whether the tail biting management tool ‘Schwanzbeiß-Interventions-Programm’ (SchwIP) had an effect on tail lesion prevalence. A total of three observers scored tail lesions from pictures of 43 328 pigs from 32 farms where SchwIP had been applied, and of 36 626 pigs from 32 control farms. Tail lesions were classified as score 0: no visible lesion, score 1: mild lesion, score 2: severe lesion and score 3: necrosis. In addition, complete loss of tail (CL) was recorded. Tail necrosis was the only tail-related carcase finding recorded by meat inspectors. (1) Tail lesion prevalences in pigs from control farms were 23.6% for score 1, 1.02% for score 2, 0.55% for score 3 and 0.41% for CL. The combined prevalence of any lesion and/or CL was 25.4%. (2) Pleurisy, lung findings, signs of inflammation in the legs, arthritis and abscesses were the most frequent meat inspection findings (prevalences of 8.46%, 8.09%, 2.99%, 0.83% and 0.23%, respectively; n=79 954 pigs). Leg inflammation, arthritis and abscesses were more prevalent in pigs with tail lesions of any degree compared with pigs without tail lesions (3.39% v. 2.83%, 1.06% v. 0.75% and 0.39% v. 0.17%, respectively; all P<0.001, n=79 954 pigs). Pigs with severe tail lesions also had more lung findings (2.00% v. 0.17%, P<0.001). (3) Tail necrosis scored during meat inspection resulted in lower prevalence than scored from pictures (0.22% v. 0.69%; n=79 954 SchwIP and control farm pigs). (4) Although tail lesion prevalence was significantly higher in pigs from SchwIP than in pigs from control farms during the first 3 months (32.2% v. 23.8%, P=0.015), it was not significantly higher during the remainder of the year (22.6 v. 26.9, 24.4 v. 21.4 and 24.0 v. 28.0, second, third and fourth quarters, respectively). In conclusion, meat inspection results in much lower tail lesion prevalences than tail lesion assessment from pictures, even if only the category ‘necrosis’ is compared. Advising farms on tail biting using the management tool SchwIP helped to decrease the prevalence of tail lesions on problem farms.

Prevalence of an inflammation and necrosis syndrome in suckling piglets

The current study describes the results obtained from clinical examination of over 4700 suckling piglets from 19 individual herds in Germany. In this cohort the prevalence of inflammation and necrosis in the tails, ears, claw coronary bands, heels and teats was determined using a pre-defined scoring system. Results show that already in the 1st days of life, piglets were affected by inflammation and necrosis of the heels (80%), claw coronary bands (50%) and tail base (20%). The praevalences of these alterations in piglets were influenced by genetics (P <0.001) and age, decreasing gradually in the 2nd week of life (P <0.001). Moreover, a correlation between tail length after tail docking and the prevalence of tail necrosis (P⩽0.04) was found. Tail and ear biting as a behavioural trait was not detected during this study. The early onset, appearance and multiple locations of clinical signs of inflammation and the positive correlation with the genetic background of the piglets may suggest an impairment of the innate immune system by infectious and non-infectious agents. This is in contrast to previously described behavioural abnormalities seen in fattening pigs. Considering the obvious reduction of animal welfare due to the described lesions, there is a need to create awareness among pig farmers and to understand the multifactorial causality involved in this inflammation and necrosis syndrome in piglets.

Role of mycotoxins in herds with and without problems with tail necrosis in neonatal pigs

This study aimed to investigate a possible involvement of mycotoxins in neonatal tail necrosis in piglets. Ten affected and 10 non-affected farms were selected. Sow feed samples were analysed for...

TIA1 is a gender-specific disease modifier of a mild mouse model of spinal muscular atrophy

Spinal muscular atrophy (SMA) is caused by deletions or mutations of Survival Motor Neuron 1 (SMN1) gene. The nearly identical SMN2 cannot compensate for SMN1 loss due to exon 7 skipping. The allele C (C+/+) mouse recapitulates a mild SMA-like phenotype and offers an ideal system to monitor the role of disease-modifying factors over a long time. T-cell-restricted intracellular antigen 1 (TIA1) regulates SMN exon 7 splicing. TIA1 is reported to be downregulated in obese patients, although it is not known if the effect is gender-specific. We show that female Tia1-knockout (Tia1−/−) mice gain significant body weight (BW) during early postnatal development. We next examined the effect of Tia1 deletion in novel C+/+/Tia1−/− mice. Underscoring the opposing effects of Tia1 deletion and low SMN level on BW gain, both C+/+ and C+/+/Tia1−/− females showed similar BW gain trajectory at all time points during our study. We observed early tail necrosis in C+/+/Tia1−/− females but not in males. We show enhanced impairment of male reproductive organ development and exacerbation of the C+/+/Tia1−/− testis transcriptome. Our findings implicate a protein factor as a gender-specific modifier of a mild mouse model of SMA.

Modified Mouse Models of Chronic Secondary Lymphedema: Tail and Hind Limb Models

Postsurgical secondary lymphedema is usually a progressive and lifelong condition lacking any curative treatment. The aim of this study was to develop new, simple surgical mouse models of chronic lymphedema, better simulating chronic nature of human postsurgical lymphedema. Two experimental mouse models of secondary lymphedema were created surgically without radiation by modifications of the previously described methods: the tail model and the hind limb model. Lymphedema formation was clinically assessed and quantitatively evaluated by measuring circumferences and limb volumes. Postmortem specimens were assessed histologically to examine the efficacy of the models. In the tail models, although a substantial frequency of tail necrosis (30.0%) was noted and the increase in circumference was maintained for only limited times postoperatively depending on the particular tail model, the overall success rate was 65.0%. In the mouse hind limb model, the overall success rate was 88.9%, and the increased circumference and limb volume were maintained over the entire study period of 8weeks. The overall success rate of the mouse hind limb model was significantly higher than that of the mouse tail model(s). We have successfully established modified mouse tail and hind limb lymphedema models via only surgical techniques without radiation, which have characteristics of chronic secondary lymphedema. The mouse hind limb model has a higher success rate than the mouse tail model and has advantages of having the healthy contralateral hind limbs as an internal control.

Gender-Specific Amelioration of SMA Phenotype upon Disruption of a Deep Intronic Structure by an Oligonucleotide

Spinal muscular atrophy (SMA), the leading genetic disease of children, is caused by low levels of survival motor neuron (SMN) protein. Here, we employ A15/283, an antisense oligonucleotide targeting a deep intronic sequence/structure, to examine the impact of restoration of SMN in a mild SMA mouse model. We show gender-specific amelioration of tail necrosis upon subcutaneous administrations of A15/283 into SMA mice at postnatal days 1 and 3. We also demonstrate that a modest increase in SMN due to early administrations of A15/283 dramatically improves testicular development and spermatogenesis. Our results reveal near total correction of expression of several genes in adult testis upon temporary increase in SMN during early postnatal development. This is the first demonstration of invivo efficacy of an antisense oligonucleotide targeting a deep intronic sequence/structure. This is also the first report of gender-specific amelioration of SMA pathology upon a modest peripheral increase of SMN.

Protein C Activation Is the Critical Thrombomodulin Function in Embryonic and Adult Survival in Mice

BACKGROUND AND APPROACH:Thrombomodulin (Thbd) is a multifunctional transmembrane molecule expressed on endothelial, hematopoietic, placental trophoblast, and various other cells. Thbd facilitates protein C and TAFI activation by thrombin, but also exerts thrombin- and protein C-independent functions to regulate complement activity and inflammatory signaling. Complete Thbd deficiency causes embryonic death through as yet unclear mechanisms, while ablation of the lectin-like Thbd domain, reduction of Thbd cofactor activity for protein C (pC) activation, or truncation of the cytoplasmic domain do not impair embryonic development1-3. Here we applied in vivo structure-function and genetic complementation analyses to differentiate between pC-dependent and -independent Thbd functions.RESULTS:Three mouse strains with various function-selective Thbd mutations were generated, i.e. ThbdPro/ΔCS, ThbdEGF4-6, and ThbdLT (Table 1). Of these, only the latter mutant with defective thrombin-binding reproduced the embryonic lethal phenotype of complete Thbd deficiency. To examine the developmental role of Thbd expression in cell types other than placental trophoblast, Thbd LoxP mice carrying a conditional Thbd allele were crossed with mice expressing Cre recombinase under control of the endogenous Meox2 gene promotor, resulting in selective ablation of Thbd in the embryo proper, but preservation in extra-embryonic placental tissue. This yielded a normal number of Thbd-deficient embryos at term pregnancy. Preservation of Thbd function in the embryonic placenta therefore was sufficient to prevent intrauterine lethality until birth. Immunohistochemistry of term embryos and adult mice (week 10-16) confirmed persistent absence of Thbd expression. Half of Thbd-null mice were lost prior to weaning (3 weeks). Surviving mice were smaller, yet exhibited normal weight gain thereafter. By 4 weeks of age, 19 of 28 Thbd-deficient animals developed grossly observable lesions, i.e. purple discoloration and swelling of tail segments, followed by, edematic swelling and discoloration of the tail and hind limb digits, distal tail necrosis and/or uni- or bilateral eye degeneration. More than half of the Thbd-null mice expired within the first 10 weeks of life, and only 2 of 28 mice survived to week 40. Neither gross anatomical inspection nor histological surveys of internal organs revealed evidence of severe thrombotic organ damage, and except for elevated IL-6, cytokine plasma levels were normal. Tamoxifen-induced Thbd LoxP gene ablation in adult mice (10-12 week old) reproduced the phenotype of Meox2Cre-Thbd-null mice. Genetic supplementation of activated PC by transgenic expression of a partially Thbd-independent murine pC zymogen variant (“hyperactivatable” protein C4), prevented perinatal lethality, partially restored normal birth weight, and largely prevented the phenotype of Thbd-null mice. However, Thbd-null females expressing the pC transgene exhibited pregnancy-induced morbidity and mortality with complete penetrance. When mated to wild type males (embryonic placenta expresses Thbd), 8 of 8 pregnancies ended in death or severe distress during gestation or postpartum.CONCLUSIONS:These findings indicate that the lack of protein C activation by Thbd-bound thrombin is the sole cause of lethality of embryonic and adult Thbd-null mice; and that Thbd expression in extra-embryonic placental tissue is sufficient to prevent intra-uterine lethality. The mortality of pregnant Thbd-null females expressing the protein C transgene constitutes a novel model for pregnancy-specific complications associated with defects in the function of natural anticoagulant pathways.REFERENCES:1. Isermann B et al. Nat Med. 2003;9(3):331-337.2. Conway EM et al. Blood. 1999;93(10):3442-3450.3. Conway EM et al. J Exp Med. 2002;196(5):565-577.4. Isermann B et al. Nat Med. 2007;13(11):1349-1358. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

EAR AND TAIL LESIONS ON CAPTIVE WHITE-TAILED DEER FAWNS (ODOCOILEUS VIRGINIANUS): A CASE STUDY.

During the 2008-2011 time period, undiagnosed lesions were observed in 21 of 150 white-tailed deer fawns (Odocoileus virginianus) that were part of a captive deer herd at Mississippi State University. Clinical findings in healthy and diseased fawns from 0 to 90 days of age included bite and scratch marks followed by moderate to severe ear and tail necrosis. Gross necropsy findings of necrotizing ulcerative dermatitis correlated with histopathologic findings that included focally severe multifocal vasculitis, vascular necrosis, and thrombosis. This article is a clinical description of these previously unreported lesions associated with tissue necrosis in young captive white-tailed deer.

In vivo and in vitro antimicrobial activity of Azadirachta indica (Lin) against Citrobacter freundii isolated from naturally infected Tilapia (Oreochromis mossambicus)

Abstract Aquaculture health management demands a constant growth in the production by stabilizing economic loss and implementing various natural plant-based bio products for treating pathogenic bacteria. Pathogenic effects of the Citrobacter freundii isolated from fingerlings of Tilapia collected from a fish farm showed symptoms like tail necrosis, septicemia, hemorrhage, and reddening of the body. Antibacterial activity of C. freundii was tested against various medicinal plants. Of the three plants screened, Azadirachta indica (Neem) showed good antibacterial activity against C. freundii. The present study revealed that A. indica plant extract at a concentration of 150 mg/l in vivo can be used as an alternative to antibiotics for treating this bacterial infection. The use of medicinal plant extract-mediated treatment was found to be very effective in controlling this bacterial infection.