Microscopic Pathologic Findings Research Articles

急性膵炎後の被包化壊死増大に伴い脾梗塞・胃壁虚血に陥り外科手術を要した1例(Acute pancreatitis and enlarged area of walled–off necrosis followed by infarction of the spleen and ischemia of the gastric wall requiring surgery: case report)

症例は40歳の男性。基礎疾患に高トリグリセリド血症があり，急性膵炎を繰り返していた。約9か月前に壊死性膵炎を発症して膵尾部と胃脾間，結腸間膜に被包化壊死を合併，以降定期的にCT画像検査による経過観察を行っていた。今回左側腹部痛を主訴に来院し，腹部CT検査において胃脾間，膵尾部に各々44mm，43mmの被包化壊死を認め，いずれも2か月前の検査時より被包化壊死の増大を認めた。またそれぞれの被包化壊死に隣接して胃穹窿部の浮腫性変化と脾動脈閉塞，脾梗塞を認めた。ヘパリン・抗菌薬・鎮痛薬による保存的治療から開始したが，発熱が継続し，炎症反応の改善も得られず，入院第4病日のCT検査では脾梗塞の状況に改善がなく，一方で被包化壊死に接する胃壁全層の虚血所見が明らかとなったため，保存的治療を断念し，緊急開腹術での膵尾部切除・脾摘出・胃部分切除術を施行した。病理所見では被包化壊死と脾動脈との間には炎症性細胞の浸潤を認め，脾動脈内腔には白色血栓を認めた。胃壁漿膜側は被包化壊死に伴う炎症所見や周囲血管内の血栓形成を認めた。被包化壊死の自然史は明確ではなく，保存的治療においては，症状や画像所見を丁寧に評価して外科的治療介入の時期を見逃さないことが重要である。 A 40–year–old male with hypertriglyceridemia and recurrent acute pancreatitis was undergoing regular imaging to monitor walled–off necrosis (WON) that had developed 9 months earlier secondary to necrotic pancreatitis. The patient presented with acute left abdominal pain. Abdominal CT showed WON by the gastrosplenic ligament (44mm) and in the pancreatic tail (43mm). Both areas grew since the last imaging 2 months earlier. Edematous changes in the fornix, splenic artery occlusion, and splenic infarction were observed adjacent to areas of WON. Despite conservative management, fever and elevated inflammatory biomarkers persisted. CT after admission showed no change in the splenic infarction. It also detected ischemia in all layers of the gastric wall adjacent to areas of WON. Emergency laparotomy was performed for pancreatic tail resection, splenectomy, and partial gastric resection. Pathologic examination showed infiltration of inflammatory cells between the areas of WON and the splenic artery and a white thrombus in the splenic artery lumen. There was inflammation and thrombus in the blood vessels surrounding areas of WON on the serosal side of the gastric wall. Although the natural history of WON is not well studied, careful evaluation of clinical findings and imaging for timely surgical intervention is required. 2012年の改訂アトランタ分類 1により膵炎の局所合併症としてみられる膵あるいは膵周囲の液体貯留は，壊死の有無と発症からの経過時間により，間質性浮腫性膵炎後に発症する急性膵周囲液体貯留（acute peripancreatic fluid collection: APFC）と壊死性膵炎後に発症する急性壊死性貯留（acute necrotic collection: ANC），それぞれが4週以上経過した膵仮性嚢胞（pancreatic pseudocyst: PPC）と被包化壊死（walled–off necrosis: WON）の4つのカテゴリーに分類された。ガイドライン 2, 3, 4ではこれら膵局所合併症に関して無症候性，非感染性のものについては保存的加療を推奨し，症候性，感染合併症例を治療適応としている。今回我々は，繰り返す急性膵炎後にWONを発症し，その経過観察中に合併した脾動脈本幹閉塞による脾梗塞と，胃後壁への炎症波及に伴う胃壁虚血に対して外科的治療を施行した1例を経験した。これまでに急性膵炎に伴う脾梗塞の発生率は約7%と報告され 5，また急性膵炎に伴い胃壁梗塞および脾梗塞を合併した報告例はある 6が，WONの自然経過における同合併症の発生に関しては明確にされていない。我々は自験例におけるWONの経時的画像所見と術中所見および切除標本の病理学的所見からその機序と治療方針について考察した。 本論文は倫理委員会の承諾を必要としない症例報告である。また，個人情報保護法に基づいて匿名化がなされており，患者本人から論文出版に関する同意を取得している。 患 者：40歳の男性 主 訴：左側腹部痛 基礎疾患：高トリグリセリド血症（LPL遺伝子異常症疑い） 生活歴：喫煙歴なし，飲酒習慣なし 現病歴：以前より急性膵炎を繰り返していた。9か月前に壊死性膵炎を罹患以降，膵尾部と胃脾間，結腸間膜にWONが形成されていた（Fig. 1a–c）。6か月前，4か月前には浮腫性膵炎を罹患し絶食，輸液，ガベキサートメシル酸塩点滴，抗菌薬，経管栄養による保存的治療を受けていた。今回来院約12時間前に突然左側腹部痛を自覚，改善を認めないため，ウォークインで救急外来を受診した。 Abdominal computed tomography (CT) with contrast (a–e: contrast–enhanced CT, delayed phase, f: contrast–enhanced CT, arterial phase). a: 9 months before admission, at the onset of WON, a solitary lesion was found in the pancreatic tail. b: 6 months before admission, cystic lesions were found in the pancreatic tail and by the gastrosplenic ligament. There was no abnormal contrast effect on the spleen. c: 4 months before admission, cystic lesions were found in the pancreatic tail and by the gastrosplenic ligament. There was no abnormal contrast effect on the spleen. d: Edematous changes in the fornix of the stomach adjacent to WON on arrival (white arrow). e: All layers of the gastric wall were poorly enhanced on day 4 of admission (black arrow). f: On CT scan at the time of admission, the splenic artery next to WON in the pancreatic tail had a contrast defect (white and black arrowheads). d–f: There was poor contrast enhancement of the spleen. 経 過：来院時のバイタルサインは意識清明，心拍数91/分，血圧169/109mmHg，呼吸数24/分，酸素飽和度97%（室内気），体温37.1℃であった。身体診察では，腹部は平坦，左側を中心に腹部全体の圧痛，筋性防御を認めた。血液検査では，白血球18,100/µL，CRP 12.15mg/dLと非特異的炎症反応上昇を認め，Fib 536mg/dL，D–dimer 1.29µg/mLと軽度の線溶系亢進を認めた。膵酵素の上昇は認めなかった（Table 1）。腹部造影CT検査（Fig. 1d）では，胃脾間に最大径44mm（前回2か月前撮影時41mm），膵尾部に最大径43mm（前回2か月前撮影時38mm）のWONを認め，膵尾部以遠の脾動脈造影欠損，脾全体の造影不良を認めた（Fig. 1f）。またWONに接した胃穹窿部壁の造影不良を認めたが（Fig. 1d白矢印），胃周囲の血管には造影欠損を認めず，胃壁の浮腫性変化と考えた。脾梗塞の診断で入院加療とし，抗菌薬，ヘパリン15,000単位/日点滴静注，疼痛管理を開始した。入院後痛みは軽減したが，発熱の持続と第3病日の血液検査では白血球17,900/µL，CRP 31.40mg/dLと炎症反応の改善を得られず，入院第4病日に腹部造影CT検査を再検したところ，胃穹隆部の壁全層に及ぶ造影欠損が明確となった（Fig. 1e黒矢印）。上部消化管内視鏡検査を施行したところ，噴門部大弯から後壁にかけて胃粘膜の虚血性変化を認めた（Fig. 2）。比較的広範囲で胃壁全層の虚血壊死が生じていると疑われたため治療方針を改め，同日，緊急開腹手術を行った。腹腔内には漿液性腹水を少量認め，大網切開し網嚢腔を開放すると胃脾間・膵尾部のWONは胃後壁・膵尾部と強固に癒着一塊となっていた。脾動脈への前面からのアプローチは炎症波及により剥離困難であったため膵脾脱転したうえで脾動脈を根部から約1cmの部位で結紮し，膵臓も同部位で切離，合わせて梗塞に陥った脾を摘出した。胃後壁はWON癒着部から1cmほど離れた部位で胃壁を貫通し，壊死した胃粘膜を視認しつつWONを含めた胃部分切除を行った。結果的に胃穹窿部から体部後壁までを切除した（手術時間3時間39分，術中出血量470mL）。病理所見では胃後壁・WON・膵尾部・脾臓は一塊となり（Fig. 3a），線維性肉芽組織からなるWONの嚢胞壁には好中球浸潤を伴う中等度の急性炎症がみられ，内部は壊死物質で満たされていた。胃壁は固有筋層に及ぶ潰瘍形成を認め，一部で胃壁全層に及ぶ凝固壊死，浮腫，高度の急性炎症を認めた。漿膜側にはWONに伴う炎症所見とともに周囲の動脈内に血栓形成を認めた（Fig. 3b）。胃壁の明らかな穿孔は認めなかった。WONと隣接する動脈との間には炎症性細胞の浸潤を認め，脾動脈内腔は白色血栓で閉塞（Fig. 3c），脾臓は凝固壊死に陥っていた。術後経過は良好で，術後4日目に経口摂取を開始，同11日目に退院した。 Upper gastrointestinal endoscopy findings. There was ischemia of the gastric mucosa from the cardia and greater curvature to the posterior wall. Macroscopic and microscopic (×20) surgical pathology findings. a: Macroscopic cystic lesions were partially solid. (b, c) Histologically, the wall of the cystic lesion consists of fibrous granulation tissue with no epithelium, acute inflammation with infiltration by neutrophils, and contained internal necrotic tissue. b: The gastric wall had ulcer formation extending to the muscularis propria, coagulative necrosis throughout all layers, edema, and severe acute inflammation. Inflammatory infiltrates and thrombus were observed in the blood vessels surrounding WON on the serosal side of the gastric wall. c: In the pancreas, there was severe fibrosis, edema, and moderate to severe acute and chronic inflammation. A white thrombus was observed in the lumen of the splenic artery. W: walled–off necrosis (WON), P: pancreas, St: stomach, Sp: spleen 急性膵炎における壊死性膵炎の発症率は10～20%とされ 7，WONは壊死性膵炎の晩期合併症として生じる。WONは膵実質および膵周囲脂肪組織の壊死部が時間とともに当初の固形から融解壊死により液状化し，周囲を肉芽性・繊維性皮膜で覆う状態になったものである。近年までWONと間質性浮腫性膵炎の晩期合併症であるPPCは明確には区別されておらず，両者を含めた検討において，経過観察症例の30～50%で重大な偶発症・合併症が発生したと報告され 8, 9，発生6週以降では自然消退は期待できず偶発症・合併症率が高いとする報告 10や，嚢胞径が6cmを超えるものでは自然消退の可能性が低下し偶発症・合併症率が高くなるとする報告 9がある。一方でManraiら 11は壊死性膵炎153名の観察研究において，143名がANCを合併し，そのうちの84名がWONに移行したが，84名のWON症例中，保存的治療を受けた23名（約27%）において局所病変の自然消失もしくは軽快が得られたと報告した。また酒井ら 12は10cmを超える巨大WONの保存的治療例を報告している。このようにWONに関する保存的治療経過の詳細は未だ不明な点もあるが，現在のガイドライン 2, 3, 4においては，膵局所合併症の種数やサイズは問わず，無症候性，非感染性の症例に対しては保存的に経過観察することが推奨され，症候性の症例では感染合併が疑われるか否か，全身状態が安定か否かにより抗菌薬投与やインターベンション治療が選択される。インターベンション治療に際しては，経皮的或いは内視鏡的経消化管的ドレナージなどの非侵襲的処置から開始し，治療効果が不十分な場合には外科的手術を行うというstep–up approach法の有用性が確認されている。 本症例はこれまでに急性膵炎を繰り返していたが，今回9か月前に発症した壊死性膵炎の際に膵尾部に造影不良域を認めたほか，結腸間膜および胃脾間に不整形脂肪織濃度上昇を認めていた。膵尾部不染領域は8か月前には最大径40mmの境界明瞭な多房性病変となり，7か月前には膵尾部病変は同サイズ，胃脾間に径50mmの嚢胞性病変，結腸間膜に径67mmの不整形脂肪織濃度上昇域を認めていた。いずれも壊死性膵炎を契機に発生しており，WONと診断し1か月ごとのCT検査を行い経過観察していた。経過中の6か月前と4か月前にはそれぞれ間質性浮腫性膵炎を発症し保存的治療を行っているが，その際はいずれの局所合併症も大きさに著変はなく，その後は2か月前のCT検査において膵尾部病変は径38mm，胃脾間病変は径41mm，結腸間膜病変は径23mmとそれぞれ縮小傾向であり，脾動脈，脾臓，胃壁の造影効果にも異常は認めなかった。したがって本症例において，脾梗塞と胃壁虚血を同時に発症したのは，膵局所合併症出現から約9か月が経過したところであった。局所合併症のサイズは続発症が増加するとされる60mmより小さい40mm前後であり，この間腹部自覚症状も認めなかったため，外来経過観察中であった。 膵局所合併症の経過中にみられる病態として，感染，消化管・胆管閉塞，腹腔内・隣接消化管や臓器への破裂・穿通，出血がある 13。本症例では胃壁虚血と脾動脈閉塞に伴う脾梗塞を発症した。膵局所合併症の消化管瘻・穿破は嚢胞性病変が消化管に癒着して線維化が進行することにより，癒着臓器の伸展が強制されて生ずると考えられる。本症例では病理所見として胃壁漿膜側にWONによる炎症波及を認め，周囲動脈内の血栓形成が散見された。炎症所見は一部固有筋層に及び，比較的広範囲の胃粘膜虚血が起き，潰瘍形成を生じたと考えられた。脾梗塞に関しては，WONによる脾動脈の物理的圧排のほか，経過中に繰り返された膵炎の炎症波及による脾動脈周囲の線維化から脾動脈狭窄を生ずる機序，血管炎に伴う血栓形成機序などの関与が考えられる。本症例は短胃動脈や胃大網動脈からの十分な側副血行路が発達する前に脾全体の梗塞を来しており，動脈周囲に好中球浸潤を伴う急性炎症所見を認めることから，徐々に進行する物理的圧迫よりも，炎症に伴って急速に進行する血栓形成の要素が大きいと考えられた。脾動脈閉塞の結果として生じた脾梗塞は，発熱，疼痛などの自覚症状が改善しない場合や，膿瘍形成，破裂，敗血症性ショック，出血などの合併症を来した場合には脾摘術の適応がある 14。梗塞範囲が増えるほど合併症のリスクが高まると考えられるが，脾機能亢進症に対して行われる部分的脾塞栓術のように，無菌性脾梗塞と適切な梗塞率が保たれていれば外科的治療を必要としないという報告 15や，胃癌術後に発症した全脾梗塞の保存的治療例の報告 16がある。本症例は当初，胃壁造影不良域を浮腫性変化と捉えていた。脾梗塞に対しては出血・感染合併による病態悪化時の手術適応を念頭に置きつつ，ヘパリン・抗菌薬・鎮痛薬による内科的治療から開始したが，発熱が続き炎症反応の改善も得られなかった。入院第4病日の再評価において脾動脈閉塞および脾梗塞の状況に変化はなく，一方で胃壁の虚血所見が明確となったため，保存的治療を断念して手術治療を行った。本症例は来院時点で左側を中心とした腹部全体の圧痛・筋性防御や非特異的炎症反応上昇という腹膜炎を疑う所見を認めており，当初からstep–up approach法に則った治療介入を検討すべきであった。本症例の術中所見においてWONは胃壁と強固に癒着していたためWONを含め一塊切除とする膵尾部切除・脾摘出・胃部分切除術を施行した。 WONの合併症治療においては，常に外科的治療に移行する選択肢を念頭に置き，症状や画像所見から臨床経過を継続的に評価することが致死的合併症を防ぐために重要である。 本症例は，繰り返す急性膵炎に生じたWONの経過観察中に脾動脈本幹閉塞からの脾梗塞および胃壁虚血を合併した。自然消退しない膵局所合併症の治療方針を検討するうえで教訓的な1例と思われ報告した。 本論文の要旨は，第49回日本救急医学会総会・学術集会において発表した。 著者全員において本症例報告に関する利益相反はない。

Important Mycosis of Wildlife: Emphasis on Etiology, Epidemiology, Diagnosis, and Pathology-A Review: PART 2.

Simple SummaryThe conservation of wildlife species is a major goal globally, due to the declining number of total populations. Fungal diseases are frequently diagnosed in veterinary practice, so knowing their characteristics, along with knowing the appropriate laboratory methods to make a correct diagnosis, are extremely important for the veterinarian. This article is the continuation of an extensive review that presents the main fungal diseases of wild animals. In this part, the second, four of these mycoses are discussed (dermatophytosis, coccidioidomycosis, blastomycosis, and sporotrichosis).Wild animals are an important component of the ecosystem, and play a major role in it. However, in recent years, there has been an astronomical increase in the incidence of wildlife mycotic diseases leading to wildlife extermination. It is important to note that most of these mycotic diseases are zoonotic, and since there is a lot of attention given to zoonosis of a bacterial or viral origin in recent times, it is important to look into the mycotic diseases which may have zoonotic potential. Previously, the authors expatiated on some major wildlife mycotic diseases. In this review, we shed light on the etiology, epidemiology, diagnosis, pathogenesis, pathogenicity, macroscopic and microscopic pathology, and hematological and serum biochemical findings of dermatophytosis, coccidioidomycosis, blastomycosis, and sporotrichosis, which are very important mycoses of wildlife.

Safety assessment of a novel, dietary pyrroloquinoline quinone disodium salt (mnemoPQQ®)

Pyrroloquinoline quinone (PQQ), a potent coenzyme antioxidant naturally occurring in foods, has been demonstrated to protect brain cells by enhancing the expression of nerve growth factors (NGF) and NGF receptors, and suppressing the fibril formation and aggression of amyloid β. We developed mnemoPQQ®, a novel PQQ disodium salt and assessed its safety in GLP compliant toxicity studies. Acute toxicity studies of mnemoPQQ® in Wistar rats revealed that its LD50 was 1825- and 1410 mg/kg body weight (bw) in male and female rats, respectively, whereas its acute dermal LD50 was >2000 mg/kg bw. mnemoPQQ® was found to be nonirritant to the skin of rabbit in an acute dermal irritation/corrosion study, and classified mnemoPQQ® as a nonirritant to the eye of rabbit in an acute eye irritation/corrosion study. Ames bacterial reverse mutation assay and in vitro Mammalian cell gene mutation test exhibited its non-mutagenic potential. In mammalian in vivo erythrocyte micronucleus test, mnemoPQQ® was classified as non-clastogenic and non-mutagenic. A 90-day sub-chronic toxicity study, conducted at and up to the highest daily dose of 600 mg/kg body weight, revealed no evidence of systemic toxicity. All rats survived the treatment without any significant abnormal clinical signs and alterations in hematology, clinical chemistry, neurological evaluation, thyroid functions, reproductive hormone levels, sperm evaluations, vaginal cytology, endocrine functions, organ weight and gross and microscopic pathology findings. No observed adverse effect level (NOAEL) of mnemoPQQ® was found to be greater than 600 mg/kg body weight. These studies affirm that mnemoPQQ® has broad spectrum safety for human consumption.

Incidental Tuberculosis in sudden, unexpected, and violent deaths in the community Lusaka, Zambia - A descriptive forensic post-mortem examination study

Tuberculosis remains a global emergency. In Zambia only 55% of tuberculosis cases are diagnosed. We performed a study to determine incidental cases of tuberculosis seen at forensic autopsy of individuals who died suddenly and unexpectedly in the community in Lusaka, Zambia. Whole-body autopsies were performed according to Standard Operating Procedures. Representative samples obtained from relevant organs were subjected to pathological examination. Information on circumstances surrounding the death was obtained. Data on patient demographics, gross and microscopic pathological findings, and cause(s) of death were analysed. Incidental tuberculosis was found in 52 cases (45 male, 7 female, age range 14-66) out of 4286 whole-body autopsies. 41/52 (80%) were aged 21-50 years. One was a 14-year old boy who died during a football match. 39/52 (75%) deaths were attributable specifically to tuberculosis only. Other deaths were due to acute alcohol intoxication(4), violence(7), ruptured ectopic pregnancy(1), bacterial meningitis (1). All the cases were from poor socio-economic backgrounds and lived in high-density areas of Lusaka. Incidental cases of active tuberculosis undiagnosed antemortem seen at forensic autopsy reflects major gaps in the national TB control programs. More investments into proactive screening, testing, treatment activities, and accurate data collection are required.

Impact of longitudinal tumor location on postoperative outcomes in patients undergoing resection for gallbladder cancer: Fundus and body vs. neck and cystic duct

It is known that gallbladder cancer (GBC) in the neck or cystic duct (NC-GBC) has a better prognosis than GBC in the fundus or body (FB-GBC), but systematic studies on this are insufficient. We performed this study to investigate the impact of longitudinal tumor location on postoperative outcomes in patients undergoing resection for GBC. A retrospective study was conducted for patients who underwent a radical resection for GBC from February 2008 to November 2017 at the Dankook University Hospital. A total of 98 patients underwent surgery for GBC, of which 77 patients who underwent curative intent surgery were included in the study. They were classified into FB-GBC and NC-GBC groups according to longitudinal tumor location, and the postoperative outcomes were compared and analyzed. There were no significant differences in the clinicopathological characteristics, TNM stage, postoperative complications, and in-hospital mortality between two groups. However, NC-GBC significantly showed more sclerotic gross type, poorer differentiation, and more lymphatic and perineural microinvasion. The radical resection rate was statistically higher in FB-GBC group (93.1% vs. 73.7%; p = 0.036) and adjuvant 5-FU based CCRT was more carried out in NC-GBC group (19.0% vs. 57.9%; p < 0.001). The recurrence rates after surgery was statistically higher in NC-GBC group (25.9% vs. 52.6%, p = 0.047), but there were no differences in disease-free survival (DFS) and overall survival (OS). Although NC-GBC showed more aggressive microscopic pathological findings and higher recurrence rate than FB-GBC, there were no differences in DFS and OS according to longitudinal tumor location of GBC.

Ocular tuberculosis in a calf.

BackgroundBovine tuberculosis is a chronic inflammatory disease that causes granuloma formation mainly in retropharyngeal, tracheobronchial, mediastinal lymph nodes and lungs of bovines. The presence of these lesions in other tissues such as the eyeball is very rare and difficult to diagnose. This study describes macroscopic and microscopic pathological findings in a calf with ocular and meningeal tuberculosis.Case presentationMarch 2019, an eight-month-old Holstein Friesian calf was identified in a dairy farm located in central Mexico with a clinical cough, anorexia, incoordination, corneal opacity and vision loss. At necropsy, pneumonia, lymphadenitis, meningitis, and granulomatous iridocyclitis were observed. The histopathological examination revealed granulomatous lesions in lung tissue, lymph nodes, meninges and eyes with the presence of acid-fast bacilli associated with Mycobacterium spp.ConclusionTo the best of our knowledge, this is the first report that describes macroscopic and microscopic pathological findings of ocular tuberculosis in cattle. This report highlights the importance of considering bovine tuberculosis in the differential diagnosis of corneal opacity and loss of vision in cattle.

No Correlation Between Presenting Symptoms Prior to Suburethral Sling Removal and Explanted Pathology Findings Suggests No Need for Routine Microscopic Pathology Evaluation.

We compared explanted midurethral sling (MUS) standard clinical pathology report findings with presenting symptoms before synthetic sling removal (SSR). An institutional review board-approved, prospectively maintained database of women who underwent a single SSR for MUS-related complication(s) was retrospectively reviewed for demographics, time interval between MUS placement and SSR, MUS type, and presenting symptoms before SSR. We compared 2 groups, one with MUS-related symptoms, including incontinence, voiding dysfunction, dyspareunia/pain, erosion, exposure, and/or urinary tract infections, and the second group characterized by self-reported symptoms attributed to the mesh material (systemic, autoimmune, or inflammatory disorder-mesh reaction group). From 2005 to 2019, 332 of 473 women met study criteria, with 24 reporting mesh reaction-related symptoms. There was no difference in microscopic pathology findings between the 2 groups. In the mesh-related group, women were younger (P = 0.041) and had more pain/dyspareunia as their primary SSR indication (P = 0.002). That group was also significantly more likely to have a preexisting diagnosis of fibromyalgia (P < 0.001) and established autoimmune disorder (P = 0.033). No significant correlation between presenting findings and sling pathology was observed. No correlation was observed when comparing pathological findings of explanted MUS with presenting symptoms of MUS-related complications, including in women with mesh reaction-related concerns.

Novel chimeric E2CD154 subunit vaccine is safe and confers long lasting protection against classical swine fever virus

E2CD154 is a vaccine candidate against classical swine fever (CSF) based on a chimeric protein composed of the E2 glycoprotein fused to porcine CD154 antigen, and formulated in the oil adjuvant Montanide™ ISA 50 V2. This vaccine confers early protection in pigs and prevents vertical transmission in pregnant sows. The objectives of this study were to assess the safety of this immunogen in piglets, to compare several doses of antigen in the formulation, and to study the duration of the immunity provided by this vaccine for up to 9 months. Three trials were conducted by immunizing pigs with a two-dose regime of the vaccine. Challenge experiments were carried out with the highly pathogenic Margarita strain. No local or systemic adverse effects were documented, and neither macroscopic nor microscopic pathological findings were observed in the vaccinated animals. The three antigen doses explored were safe and induced CSF protective neutralizing antibodies. The dose of 50 μg was selected for further development because it provided the best clinical and virological protection. Finally, this protective immunity was sustained for at least 9 months. This study demonstrates that E2CD154 vaccine is safe; defines a vaccine dose of 50 μg antigen, and evidences the capacity of this vaccine to confer long term protection from CSFV infection for up to 9 months post- vaccination. These findings complement previous data on the evaluation of this vaccine candidate, and suggest that E2CD154 is a promising alternative to modified live vaccines in CSF endemic areas.

DNA Vaccine Treatment in Dogs Experimentally Infected with Trypanosoma cruzi.

Chagas disease is a chronic and potentially lethal disorder caused by the parasite Trypanosoma cruzi, and an effective treatment has not been developed for chronic Chagas disease. The objective of this study was to determine the effectiveness of a therapeutic DNA vaccine containing T. cruzi genes in dogs with experimentally induced Chagas disease through clinical, pathological, and immunological analyses. Infection of Beagle dogs with the H8 T. cruzi strain was performed intraperitoneally with 3500 metacyclic trypomastigotes/kg body weight. Two weeks after infection, plasmid DNA immunotherapy was administered thrice at 15-day intervals. The clinical (physical and cabinet studies), immunological (antibody and cytokine profiles and lymphoproliferation), and macro- and microscopic pathological findings were described. A significant increase in IgG and cell proliferation was recorded after immunotherapy, and the highest stimulation index (3.02) was observed in dogs treated with the pBCSSP4 plasmid. The second treatment with both plasmids induced an increase in IL-1, and the third treatment with the pBCSSP4 plasmid induced an increase in IL-6. The pBCSP plasmid had a good Th1 response regulated by high levels of IFN-gamma and TNF-alpha, whereas the combination of the two plasmids did not have a synergistic effect. Electrocardiographic studies registered lower abnormalities and the lowest number of individuals with abnormalities in each group treated with the therapeutic vaccine. Echocardiograms showed that the pBCSSP4 plasmid immunotherapy preserved cardiac structure and function to a greater extent and prevented cardiomegaly. The two plasmids alone controlled the infection moderately by a reduction in the inflammatory infiltrates in heart tissue. The immunotherapy was able to reduce the magnitude of cardiac lesions and modulate the cellular immune response; the pBCSP treatment showed a clear Th1 response; and pBCSSP4 induced a balanced Th1/Th2 immune response that prevented severe cardiac involvement. The pBCSSP4 plasmid had a better effect on most of the parameters evaluated in this study; therefore, this plasmid can be considered an optional treatment against Chagas disease in naturally infected dogs.

Lung microwave ablation – an in vivo swine tumor model experiment to evaluate ablation zones

Purpose To evaluate microwave ablation (MWA) algorithms, comparing pulsed and continuous mode in an in vivo lung tumor mimic model Materials and methods A total of 43 lung tumor-mimic models of 1, 2 or 3 cm were created in 11 pigs through an intra-pulmonary injection of contrast-enriched minced muscle. Tumors were ablated under fluoroscopic and 3D-CBCT-guidance using a single microwave antenna. Continuous (CM) and pulsed mode (PM) were used. According to tumor size, 3 different algorithms for both continuous and pulsed mode were used. The ablation zones were measured using post-procedural 3D-CBCT and on pathologic specimens. Results Two radiologists measured the ablation zones on CBCT and they significantly correlated with macroscopic and microscopic pathological findings: r = 0.75 and 0.74 respectively (p < 0.0001) (inter-observer correlation r = 0.9). For 1, 2 and 3 cm tumors mimics lesions (TMLs), mean maximal and transverse ablation diameters were 3.6 0.3 × 2.2 0.3 cm; 4.1 0.5 × 2.6 0.3 cm and 4.8 0.3 × 3.2 0.3 cm respectively using CM; And, 3.0 0.2 × 2.1 0.2 cm; 4.0 0.4 × 2.7 0.4 cm and 4.6 0.4 × 3.2 0.4 cm respectively for PM, without any significant difference except for 1 cm TMLs treated by PM ablation which were significantly smaller (p = 0.009) The sphericity index was 1.6, 1.6, 1.5 and 1.4, 1.5, 1.4 at 1, 2 and 3 cm for CM and PM respectively, p = 0.07, 0.14 and 0.13 for 1, 2 and 3 cm tumors mimics. Conclusion Microwave ablation for 1–3 cm lung tumors were successfully realized but with a moderate reproducibility rate, using either CM or PM. Immediate post ablation CBCT can accurately evaluate ablation zones.

Morphological and histopathological changes in placentas of pregnancies with intrauterine growth restriction.

Aim: The definition of fetal growth restriction (FGR) refers to the incapability of a fetus to achieve the appropriate estimated growth, with expected fetal weight below the 10th percentile calculated for its gestational age. Placental factors and hypoxemia are considered to be essential elements with influence on intrauterine growth restriction (IUGR) and fetal death. The purpose of the present study was to investigate the macroscopic and microscopic pathological findings regarding the placentas in pregnancies complicated by influence on IUGR. Patients, Materials and Methods: Our study included 42 third-trimester pregnant patients admitted to the Cuza Vodă Hospital of Obstetrics and Gynecology, Iaşi, Romania, in the last three years. Soon after delivery, the 42 placentas were collected and analyzed; 32 placentas came from cases previously diagnosed with influence on IUGR and were included in our study group. Ten other placentas included in the control group were selected from uncomplicated pregnancies. Standard Hematoxylin–Eosin (HE) staining method, as well as Periodic Acid–Schiff (PAS) staining, and immunohistochemical techniques for cluster of differentiation 31 (CD31) and collagen IV were used in order to highlight the morphological features of the studied placentas. Results: Our study revealed that reduced placental dimensions and eccentric umbilical cord insertion are correlated with the birthweight of the fetuses with IUGR (p<0.05). The most common histological finding in our study group was placental infarction later correlated with IUGR, but a certain causality could not be demonstrated, as this finding was also present in normal pregnancies. Other histopathological findings were also present in the influence on IUGR group, such as fibrin deposits, diffuse calcification, chronic villitis, avascular chronical villi, with no significant statistical correlations. CD31 was strongly immunoexpressed in the villous endothelial cells. Collagen IV presented a strong immunoreaction in the basement membrane and mesenchyme of the placental villi. Conclusions: Our study revealed a correlation between the dimensions of the diameters and volume of the maternal placenta and the presence of influence on IUGR. Moreover, it confirms the available data suggesting that the place of insertion of the umbilical cord is correlated with the weight of the fetus. Further studies with extended panel antibodies are needed in order to determine and complete the role of these morphological changes in the development of influence on IUGR.

Systemic tuberculosis by MYCOBACTERIUM BOVIS in a free-ranging MARSICAN brown bear (URSUS ARCTOS MARSICANUS): a Case report

BackgroundMycobacterium bovis is known to have a wide host range and has been isolated from numerous free-ranging wildlife species, carnivores included. In bears, M. bovis has been previously reported only from a culture of pooled lymph nodes of a black bear (Ursus americanus) in the absence of lesions. The aims of this study were to describe gross and microscopic pathological findings of M. bovis tuberculosis in a deceased Marsican brown bear (Ursus arctos marsicanus).Case presentationIn March 2014, an adult female Marsican brown bear was found in the Abruzzo, Lazio and Molise National Park (Italy) showing severe non-specific clinical signs. The animal died soon after its discovery and the carcass was submitted to post-mortem examination to identify the cause of death. The bear was diagnosed with a severe Mycobacterium bovis infection, with both pathological and microbiological aspects suggesting ongoing generalization. A presumptive diagnosis of mycobacterial infection was initially made based on gross findings. Histopathology showed the presence of acid-fast bacilli in all sampled tissues along with poorly organized granulomatous lesions. Slow-growing Mycobacterium sp. was isolated from multiple organs (intestine, mesenteric lymph nodes, liver, spleen, lung and kidneys). The PCR and sequencing algorithm identified the Mycobacterium sp. isolate as M. bovis. Spoligotyping demonstrated that the M. bovis isolate belonged to spoligotype SB0120.ConclusionsThis is the first report of lethal M. bovis tuberculosis infection in a free-ranging brown bear. This pathogen could have serious adverse effects in an endangered relic population such as the Marsican brown bear. Stricter application of health regulations in force, surveillance of M. bovis infections in wild ungulates and carnivore scavengers, along with dismissal of supplementary feeding points intended for cattle or wildlife, are warranted to control the presence of bovine tuberculosis in wild and domestic animals in protected areas.

Clinico-pathological evaluation of non-neoplastic and neoplastic skin lesions: A study of 100 cases

Clinico-pathological evaluation of non-neoplastic and neoplastic skin lesions: A study of 100 cases - IJPO- Print ISSN No: - 2394-6784 Online ISSN No:- 2394-6792 Article DOI No:- 10.18231/2394-6792.2019.0021, Indian Journal of Pathology and Oncology-Indian J Pathol Oncol

Leiomioma del pezón en un varón: presentación de un caso

Leiomyoma of the nipple is a rare benign tumour that is thought to arise from smooth muscle fibres in the subareolar tissue of the breast. We report the case of a male patient with a leiomyoma arising from his left nipple-areola complex. The clinical characteristics, gross and microscopic pathologic findings, and management of this lesion are discussed.

A144 - A Comparison of Pre-operative Endoscopic Biopsy Findings to Gastric Specimens after Sleeve Gastrectomy

Laparoscopic sleeve gastrectomy (LSG) as a primary weight loss procedure has increased in popularity and is now the most frequently performed bariatric operation for weight loss in the United States. As part of the procedure, the resected portion of the stomach is routinely sent to pathology for analysis. In a program that routinely performs pre-operative endoscopy with biopsies on all patients; we question the utility of sending all specimens to pathology after surgery. We hypothesized that there would be a high concordance of microscopic pathological findings between endoscopic and surgical specimens, and therefore pathologic examination of all post-surgical specimens may be redundant and unnecessarily increase costs.

Urinary mulberry cells and mulberry bodies are useful tool to detect late-onset Fabry disease.

Fabry disease is an X-linked lysosomal storage disorder caused by a lack of α-galactosidase A activity, which leads to the accumulation of globotriaosylceramide in various organs. A complete lack of α-galactosidase A activity in a hemizygous male is the classical phenotype, and some hemizygous males show primarily cardiac and/or renal symptoms that appear in adulthood; this is called the variant type or the late-onset type. The kidney and heart are the major target organs, with damage to these organs related to mortality. Thus, in Fabry patients, early detection and early treatment are critical to longevity. Here, we present a 55-year-old Japanese male patient who was diagnosed with late-onset Fabry nephropathy with cardiomyopathy but with no abnormal urinary findings except for urinary mulberry cells and mulberry bodies. In spite of the absence of abnormal urinary findings, the light microscopic and electron microscopic pathological findings showed extensive deposition of globotriaosylceramide to podocytes. In this paper, we propose that the presence of mulberry cells and mulberry bodies can be used for the earlier detection of Fabry nephropathy, especially the late-onset type.

Pathology of Mouse Models of Accelerated Aging.

Progeroid mouse models display phenotypes in multiple organ systems that suggest premature aging and resemble features of natural aging of both mice and humans. The prospect of a significant increase in the global elderly population within the next decades has led to the emergence of "geroscience," which aims at elucidating the molecular mechanisms involved in aging. Progeroid mouse models are frequently used in geroscience as they provide insight into the molecular mechanisms that are involved in the highly complex process of natural aging. This review provides an overview of the most commonly reported nonneoplastic macroscopic and microscopic pathologic findings in progeroid mouse models (eg, osteoporosis, osteoarthritis, degenerative joint disease, intervertebral disc degeneration, kyphosis, sarcopenia, cutaneous atrophy, wound healing, hair loss, alopecia, lymphoid atrophy, cataract, corneal endothelial dystrophy, retinal degenerative diseases, and vascular remodeling). Furthermore, several shortcomings in pathologic analysis and descriptions of these models are discussed. Progeroid mouse models are valuable models for aging, but thorough knowledge of both the mouse strain background and the progeria-related phenotype is required to guide interpretation and translation of the pathology data.

Dermatotoxicology

The minipig has long been identified as the nonrodent animal of choice for preclinical safety evaluation of topically applied materials. This article reviews types of topical applications, study designs, and practical considerations. Dermal administration to the minipig presents multiple challenges. Issues to be considered are the area of exposure, the nature of the test article, the need for covering of the dose site, preparation of the dose site, and procedures for administration. The potential for cross-contamination (exposure of control animals to test article) is very high in topical studies and appropriate safeguards to prevent this are discussed. Topical administration to the intact skin is the most commonly utilized form of safety evaluation, but procedures have also been developed for intradermal administration and for the use of nonintact/wounded skin; these will be discussed. Evaluation of local (topical) effects is critical and can become complicated. Considerations in evaluations of the skin are interobserver variability, use of different scoring systems and discrepancies betweenin vivoobservations, necropsy observations, and microscopic pathology findings. Causes for apparent discrepancies and suggestions for practical resolution through appropriate procedures are discussed. Practical issues in necropsy and histotechnology procedures and techniques to optimize preservation of skin are also discussed.

Placental Mesenchymal Dysplasia

AbstractThis is an updated review of the recently described entity, placental mesenchymal dysplasia, which has been shown to have recognizable antenatal characteristics, distinctive gross and microscopic pathologic findings, associated fetal and neonatal morbidity and mortality, and unique pathogenic mechanisms. Recent understanding of the frequently associated genotype, androgenetic biparental mosaicism, is reviewed and the spectrum of changes discussed.

Abstract 641: Trastuzumab-dolaflexin, a highly potent Fleximer-based antibody-drug conjugate, demonstrates a favorable therapeutic index in exploratory toxicology studies in multiple species

Abstract Antibody-drug conjugates (ADCs) represent a promising drug class that relies on monoclonal antibody recognition of specific cancer-associated antigens for targeted delivery of chemotherapeutic agents. Fleximer® ADCs utilize a polymer-based conjugation platform to enable high drug-antibody ratios (DAR) and significantly greater anti-tumor potency compared to ADCs with DAR 3-4. We previously showed that trastuzumab-dolaflexin (T-dolaflexin), a HER2-targeted ADC comprised of the antibody trastuzumab conjugated to ∼15 proprietary auristatin molecules via Fleximer, has excellent pharmacokinetics and efficacy in mouse models (AACR 2014 Abstract #2645). T-dolaflexin is efficacious at a single dose of 0.67 mg/kg in mouse xenograft models, and achieves prolonged tumor-free survival after a single 2 mg/kg dose in a low HER2-expressing model that is insensitive to ado-trastuzumab emtansine (T-DM1). We sought to test the tolerability of T-dolaflexin in exploratory mouse, rat and non-human primate toxicology studies. Cohorts of 6 mice were treated with a single dose of vehicle or T-dolaflexin at 20, 30 or 40 mg/kg and followed for 21 days. The 20 and 30 mg/kg doses were well-tolerated based on body weight loss and mortality and achieved a therapeutic index (TI) in mouse &amp;gt;40X. Cohorts of 4 female cynomolgus monkeys were treated with a single dose of vehicle or T-dolaflexin at 0.67, 1.34 or 2.68 mg/kg (payload doses 600, 1200 and 2400 μg/m2, respectively). Day 3 necropsy was performed on 2 animals per cohort, with recovery necropsy on Day 22 in the remaining 2 animals. All animals survived until scheduled necropsy with limited body weight loss. There were no test-article related findings on gross pathology. Most notable clinical pathology findings were transaminase elevations (primarily AST), and decreased platelet counts at Day 8. One high dose animal demonstrated laboratory findings consistent with hepatic injury with full recovery by Day 22 and no findings on gross or microscopic pathology at necropsy. There was no evidence of myelosuppression. Microscopic pathology findings were limited, with no test-article related findings in HER2-expressing organs including heart, lungs and GI tract. All doses were considered well-tolerated. Toxicokinetics demonstrated good stability of drug conjugate in plasma with t1/2 ∼5 days (comparable to antibody t1/2) and minimal exposure to free payload. Plasma exposure at the 2.68 mg/kg dose in cyno was several fold higher than mouse exposure at the minimally efficacious dose of 0.67 mg/kg, and was comparable to the mouse exposure associated with prolonged tumor free survival after a single 2mg/kg dose. Trastuzumab-dolaflexin, a highly potent Fleximer-based ADC, demonstrates a favorable TI in tumor models with low HER2 expression where current HER2-directed therapies are inactive. Citation Format: Natalya Bodyak, Alex Yurkovetskiy, Peter U. Park, Dmitry R. Gumerov, Michael DeVit, Mao Yin, Joshua D. Thomas, LiuLiang Qin, Timothy B. Lowinger, Donald A. Bergstrom. Trastuzumab-dolaflexin, a highly potent Fleximer-based antibody-drug conjugate, demonstrates a favorable therapeutic index in exploratory toxicology studies in multiple species. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 641. doi:10.1158/1538-7445.AM2015-641