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Despite fast advances in genomics and proteomics, monoclonal antibodies (mAbs) are still a valuable tool for areas such as the evolution of basic research in stem cells and cancer, for immunophenotyping cell populations, diagnosing and prognosis of diseases, and for immunotherapy. To summarize different subtractive immunization approaches successfully used for the production of highly specific antibodies, we identified scientific articles in NCBI PubMed using the following search terms: subtractive immunization, monoclonal antibody, tolerization, neonatal, high-zone tolerance, masking immunization. Patent records were also consulted. From the list of results, we included all available reports, from 1985 to present, that used any enhanced immunization technique to produce either polyclonal or monoclonal antibodies. Our examination yielded direct evidence that these enhanced immunization techniques are efficient in obtaining specific antibodies to rare epitopes, with different applications, such as to identify food contaminants or tumor cells.

Background Overexpression of CXC chemokine receptor 4 (CXCR4+) is a poor prognostic factor in adult acute myeloid leukemia (AML); however, its prognostic significance in pediatric AML is unclear. Procedure This retrospective study examined the prognostic significance of CXCR4+ in pediatric AML patients enrolled in the Japanese Pediatric Leukemia/Lymphoma Study Group AML-05 study. Results In the total cohort (n = 248), no significant differences were observed between CXCR4+ patients (n = 81) and CXCR4− patients (n = 167) in terms of 3-year overall survival (OS) (69.4% vs. 75.2%, P = 0.44). However, there was a significant difference in 3-year OS between CXCR4+ and CXCR4− patients in the low-risk (LR) group (n = 93; 79.2% vs. 98.3%, P = 0.007). CXCR4+ patients in the t(8;21) AML without KIT mutation group had a significantly worse 3-year OS than CXCR4− patients (n = 44; 76.1% vs. 100.0%, P = 0.01). Multivariate Cox regression analysis identified CXCR4+ as a poor prognostic factor for OS in LR AML patients (hazard ratio, 11.47; P = 0.01). Consistent with the data for survival analysis, CXCR4+ patients in the t(8;21) AML group had a higher incidence of splenomegaly than CXCR4− patients (25.9% vs. 5.9%, P = 0.03). Conclusions These results suggest that CXCR4+ is a poor prognostic factor for LR patients, particularly t(8;21) patients without KIT mutation. The poor outcome was only applicable to OS, not relapse-free survival (RFS); thus, CXCR4+ may be associated with a poor prognosis after recurrence. Intensive therapy, including administration of CXCR4 antagonists, may be promising for pediatric AML patients with LR.

There is increasing recognition of the role of Toll-like receptor 3 (TLR3) in noninfectious inflammatory diseases, but the function of TLR3 in inflammatory skin diseases is unclear. We investigated the functions of TLR3 in allergic and irritant contact dermatitis (ICD). The contact hypersensitivity (CHS) response was lower in Toll-like receptor 3 knockout (Tlr3 KO) mice, and was greater in TLR3 transgenic (Tg) mice than in wild-type (WT) mice after challenge with 2,4,6-trinitro-1-chlorobenzene. Adoptive transfer of immunized lymph node cells from Tlr3 KO mice induced CHS in WT recipients. In contrast, adoptive transfer of those from WT mice did not fully induce CHS in Tlr3 KO recipients. The ICD reaction following croton oil application was lower in Tlr3 KO mice, and was greater in TLR3 Tg mice than in WT mice. Maturation, migration, and antigen presentation of dendritic cells and proliferation of lymphocytes between WT mice and Tlr3 KO mice were comparable. These results show that TLR3 enhances antigen-independent skin inflammation in the elicitation phase of allergic contact dermatitis and in ICD.

Raw fish consumption is increasing worldwide. Since around the year 2000, western regions of Japan have reported a foodborne disease of unknown cause that occurred after the consumption of flounder. In October 2010, a particularly large outbreak was reported in these regions among individuals who consumed flounder fish that had been raised in aquaculture systems. The median incubation period was 5 h (range, 4-19 h), and the most frequently reported symptom was diarrhea (80%). The risk estimate of the consumption of flounder was significantly higher than that of the development of symptoms (odds ratio=9.50; 95% confidence interval, 1.59-∞). According to a trace-back investigation, all of the flounder responsible for the outbreak were raised in aquaculture systems. Microscopic examination revealed that the median amount of Kudoa septempunctata present in the muscle of flounder fish from the aquaculture farm was 4.5×10(3) spores/g (range, 1.0×10(3)-9.6×10(6) spores/g). The number of K. septempunctata spores required for the development of illness, as estimated using the Monte Carlo simulation, was 7.2×10(7) spores/g; therefore, thus this might be the minimum ingestion threshold for the development of gastrointestinal symptoms. As a public health measure, the current study results should be referred to for the prevention of the gastrointestinal symptoms related to the consumption of flounder; the national public health authority has disseminated these results. We concluded that K. septempunctata-contaminated flounder fish were associated with the gastrointestinal symptoms of this recent outbreak.

Background Accumulating evidence suggests that Toll-like receptor (TLR)-3 signaling is involved in non-infectious immune and inflammatory reactions as well as in viral infections. The skin of patients with atopic dermatitis (AD) is often infected with virus and bacteria, leading to the aggravation of atopic symptoms. These findings suggest TLR3 signaling may be involved in the pathogenesis of AD, but the exact role of TLR3 in AD remains to be defined. Objective The purpose of this study was to investigate the role of TLR3 in chronic contact hypersensitivity reactions induced by repeated elicitation, resembling the features of AD. Methods Wild-type (WT) and Toll-like receptor 3 knockout (Tlr3 KO) mice were sensitized, and chronic contact hypersensitivity reactions were elicited in their ear skin via repeated application of a hapten, 2,4,6-trinitro-1-chlorobenzene (TNCB) or oxazolone. Results The Tlr3 KO mice exhibited less ear swelling, less leukocyte infiltration into the skin, and lower serum total IgE levels than WT mice after hapten challenge. The Tlr3 KO mice also displayed lower expression levels of inflammatory cytokines (interleukin (IL)-33, IL-4, IL-10, and interferon-ɤ in their TNCB-treated ear skin than WT mice. Conclusion These results showed that TLR3 deficiency suppressed the development of chronic contact hypersensitivity reactions, suggesting that TLR3 signaling may participate in the pathogenesis of AD.

Kindler syndrome is a rare autosomal recessive genodermatosis characterized by trauma-induced acral blisters in infancy and childhood, photosensitivity, and progressive poikiloderma. Other clinical features include chronic erosive gingivitis, dysphagia, esophageal and urethral strictures, ectropion, and an increased risk of mucocutaneous squamous cell carcinoma. We describe a patient with Kindler syndrome associated with squamous cell carcinoma of the skin and larynx. He had squamous cell carcinoma on his left knee with simultaneous unresectable laryngeal carcinoma at the age of 43 years. The squamous cell carcinoma on his knee was excised and the laryngeal carcinoma was treated with radiation therapy. Although pathophysiology of Kindler syndrome and its frequency of association with cancer are still not fully elucidated, we speculate that long-term erosion and regeneration of mucosal and cutaneous surfaces may have induced squamous cell carcinoma on the patient’s knee and larynx.

Interstitial granulomatous dermatitis (IGD) is a reactive phenomenon accompanying disorders including autoimmune disease, lymphoproliferative disorders and drug reactions. Histologically, IGD shows a granulomatous infiltrate surrounding piecemeal fragmentation of collagen in the diffuse interstitium. IGD presents with linear cords, papules and plaques mainly on the trunk and extremities. Herein, we report a case with peculiar clinical features that were histopathologically consistent with IGD. A 74-year-old man presented with periungual painful erythema, nail deformity of all fingers and labial, penile and anal erosive erythema. Histopathological examination of the lesions showed interface dermatitis and a diffuse interstitial granulomatous infiltrate mainly composed of CD68-positive histiocytes and lymphocytes. Degenerative collagen bundles were also observed in granulomas. C-reactive protein and the white blood cell count were elevated, but further examinations did not reveal systemic inflammatory disorders such as autoimmune disease, lymphoproliferative disorder, inflammatory bowel disease or drug hypersensitivity. The lesions were successfully treated with oral and topical steroids.

日本におけるICU入室患者家族のニーズの構造を明確にすることを目的とし,家族を対象として質問紙調査を実施した。112名の回答を分析した結果,すべての家族に共通して「必要・重要」とされる項目と,家族によって回答に散らばりのみられる項目があり,家族のニーズは二層構造となっていることが示唆された。すべての家族から「必要・重要」という回答を得た15項目についてKJ (Kawakita Jiro)法を実施した結果,「医療技術・スタッフへの期待や信頼」10項目と「病状・治療に関する基本的な情報」5項目に分類された。また回答に散らばりのみられた33項目について探索的因子分析を行い,抽出された3因子についてそれぞれステップワイズ探索的因子分析をした結果,「医療スタッフからのサポートに関するニーズ」,「情報を適時に得たいというニーズ」,「面会における融通性に関するニーズ」の3因子15項目からなるニーズ尺度が作成された。本尺度については一部の信頼性と妥当性が実証され,ICU入室患者家族のニーズの個人差を測定するのに適切な尺度であるといえた。

Patients with atopic dermatitis are usually responsive to conventional treatment such as topical steroids; however, they are sometimes refractory to the treatment. The influence of contact sensitivities on the course of patients with recalcitrant atopic dermatitis is not known. The aim of this study was to investigate whether contact sensitivities affect the course of patients with recalcitrant atopic dermatitis. We evaluated 45 patients with atopic dermatitis who had failed conventional therapy. Patch testing was performed with the Japanese standard series, metal series and/or suspected items. A total of 15 patients had a positive patch test reaction to at least one allergen. The most common allergens were nickel, topical drugs and rubber accelerators. Avoidance of products or food containing allergic substances greatly or partially improved skin symptoms in nine patients. These results suggest that contact allergens and metals may be critical factors causing eczematous lesions in patients with recalcitrant atopic dermatitis.

肝癌と胃癌は重複癌の中では比較的多い組み合せであるが, 両者とも根治手術可能であった症例は少ない.今回われわれは, 細小肝癌と早期胃癌の重複癌を同時に切除しえた症例を経験したので報告する.症例は56歳男性で心窩部異和感を訴え受診した.胃内視鏡にてIIc型早期胃癌, さらに腹部超音波, 腹部computed tomographyおよび腹部血管造影にて細小肝癌と診断, 胃亜全摘術および肝部分切除術を同時に施行した.術後1年で肝癌再発を認めたため, 経大腿動脈灘て固有肝動脈内にカテーテルを挿管し動注癌化学療法を施行, 術後1年4か月の現在, 健在である.本邦において根治手術が可能であった肝胃重複癌の報告は本症例を含めて27例ある.これらの症例を検討すると予後は肝硬変, 肝癌により左右されており, 肝癌の早期発見とともに肝癌に対する積極的な集学的癌治療力が必要であると考えられた.なお, 細小肝癌と早期胃癌の組給わせは本症例が3例目である.

Background: The outcome of acute myeloid leukemia (AML) has been improved by advanced risk-stratified chemotherapy based on prognostic factors. However, the long-term survival rate remains 60-70% and further investigation is needed. Core binding factor (CBF)-AML, characterized by t(8;21) or inv(16)/t(16;16), is the most frequent subtype in pediatric AML. Although CBF-AML is generally classified as a low-risk (LR) group, approximately 30% of the patients relapsed within 3 years in the Japanese Pediatric Leukemia/Lymphoma Study Group (JPLSG) AML-05 trial. This finding suggests that the LR patients had other risk factors that could account for their adverse outcome. CXC chemokine receptor 4 (CXCR4) is a G protein-coupled receptor that helps leukemia cells adhere to bone marrow stromal cells that affect the cell survival and proliferation. There are several reports that CXCR4 overexpression is an adverse prognostic factor in adult AML; however, few studies have examined their significance in pediatric AML. Here, we retrospectively examined whether CXCR4 overexpression can be an adverse prognostic factor in pediatric AML with LR. Patients and Methods: From 2006 to 2010, 485 de novo pediatric AML patients aged Results and Discussion: A total of 248 samples were available for this study. No significant differences in the clinical outcomes were observed between these 248 patients and the 195 patients who did not have CXCR4 data (EFS, P =0.55; OS, P =0.87). CXCR4 overexpression (CXCR4 +) was present in 81 patients (32.7%). CXCR4 + was not correlated with age, sex, and WBC count. CBF-AML was similarly distributed between CXCR4 + and CXCR4 - patients. The frequency of FLT3 -ITD in CXCR4 + patients was significantly higher in CXCR4 - patients (P =0.049), while there were no significant differences in that of KIT and CEBPA mutation (P =0.08 and 0.71, respectively). In total (n=248), no significant differences were observed between CXCR4 + and CXCR4 - patients in 3-year OS (68.6% vs. 75.1%, P =0.42) and 3-year EFS (65.6% vs. 58.9%, P =0.27). However, in the LR group (n=93), a significant difference was observed between CXCR4 + and CXCR4 - patients in 3-year OS (79.2% vs. 98.3%, P =0.008), whereas no significant difference was observed in 3-year EFS (83.3% vs. 69.6%, P =0.33). We have revealed that KIT mutations are adverse prognostic factors in t(8;21) pediatric AML in our recent study (Leukemia, 2015). In t(8;21) AML without KIT mutations (n=44), a significant difference was observed between CXCR4 + and CXCR4 - patients in 3-year OS (76.1% vs. 100.0%, P =0.01), whereas no significant difference was observed in 3-year EFS (77.8% vs. 74.2%, P =0.85). Multivariate Cox regression analysis, including CXCR4 +, KIT mutation, and age identified CXCR4 + as the only prognostic factor predicting poor OS in the cohort of AML with LR (hazard ratio, 11.01; P =0.01), but not EFS (hazard ratio, 0.54; P =0.21). The results suggest that CXCR4 + is associated with an adverse prognosis in pediatric AML with LR. The adverse outcome was restricted in OS, not in EFS, suggesting that CXCR4 + may be associated with the poor prognosis after recurrence, although the underlying mechanism remains unknown. Furthermore, it is noteworthy that the OS of LR patients with CXCR4 - was quite high, which should be confirmed in larger studies. Conclusions: We identified the adverse prognostic significance of CXCR4 overexpression in pediatric AML with LR. The adverse prognosis may be restricted after recurrence. Further studies of the underlying biology are required. There are several promising CXCR4 antagonists including AMD3100, therefore, our findings will help to develop better therapeutic approaches in pediatric AML with LR. Figure 1. Figure 1. Disclosures No relevant conflicts of interest to declare.

Changes in the cardiac sarcolemma in myocardial infarction were studied by both determination of Na+-K+-ATPase activity and SDS gel electrophoretic analysis of sarcolemmal proteins in the canine heart. Nlnety minutes after coronary ligation. Na+-K+-ATPase activity in ischemic myocardium was decreased significantly to approximately 36% of that of non-ischemic myocardium, and it remained at the lower level for 28 days. By SDS gel electrophoresis, reduction of the protein band with molecular weight of 111.000, which is suggestive of the main component of ATPase, was observed simultaneously with the reduction of Na+-K+-ATPase activity. These results indicate that ischemia for 90 minutes produces substructural changes in the sarcolemma indicating irreversible myocardial changes.

Background Interleukin (IL)-21 is a member of the type I cytokine family and plays a role in the pathogenesis of T helper type 2 allergic diseases. It has been reported that IL-21 expression is upregulated in acute skin lesions in atopic dermatitis (AD) patients; however, little is known about the serum IL-21 levels of AD patients. The aim of this study was to quantify the serum IL-21 levels of AD patients and to evaluate the relationships between the serum IL-21 level and disease severity, laboratory markers, and eruption type in AD patients. Methods We measured the serum IL-21 levels of adult AD patients and healthy control subjects using an enzyme-linked immunosorbent assay. Results The adult AD patients exhibited significantly higher serum IL-21 levels than the healthy control subjects. A comparison of the patients' serum IL-21 levels based on the clinical severity of their AD revealed that the patients with severe AD demonstrated significantly higher serum IL-21 levels than those with mild AD and the healthy control subjects. The serum IL-21 levels were significantly correlated with the skin severity score, and especially with the degree of acute lesions such as erythema and edema/papules. The serum IL-21 level was not associated with laboratory markers, such as the serum IgE level, the serum thymus and activation-related chemokine level, blood eosinophilia, and the serum lactate dehydrogenase level. Conclusions These results suggest that IL-21 might be involved in the pathogenesis of AD, especially the development of acute skin lesions.