Characterisation of the T-cell response to Ebola virus glycoprotein amongst survivors of the 2013-16 West Africa epidemic.

Zaire ebolavirus (EBOV) is a highly pathogenic filovirus which can result in Ebola virus disease (EVD); a serious medical condition that presents as flu like symptoms but then often leads to more serious or fatal outcomes. The 2013-16 West Africa epidemic saw an unparalleled number of cases. Here we show characterisation and identification of T cell epitopes in surviving patients from Guinea to the EBOV glycoprotein. We perform interferon gamma (IFNγ) ELISpot using a glycoprotein peptide library to identify T cell epitopes and determine the CD4+ or CD8+ T cell component response. Additionally, we generate data on the T cell phenotype and measure polyfunctional cytokine secretion by these antigen specific cells. We show candidate peptides able to elicit a T cell response in EBOV survivors and provide inferred human leukocyte antigen (HLA) allele restriction. This data informs on the long-term T cell response to Ebola virus disease and highlights potentially important immunodominant peptides.

Nanogel-based nasal ghrelin vaccine prevents obesity.

Obesity is associated with multiple comorbidities such as cardiovascular diseases and has a huge economic impact on the health-care system. However, the treatment of obesity remains insufficient in terms of efficacy, tolerability, and safety. Here we created a nasal vaccine against obesity for the first time. To avoid the injectable administration-caused pain and skin-related adverse event, we focused on the intranasal route of antigen delivery. We developed a vaccine antigen (ghrelin-PspA (pneumococcal surface protein A)), which is a recombinant fusion protein incorporating ghrelin, a hormone that stimulates food intake and decreases energy expenditure, and PspA, a candidate of pneumococcal vaccine as a carrier protein. Ghrelin-PspA antigen was mixed with cyclic di-GMP adjuvant to enhance the immunogenicity and incorporated within a nanometer-sized hydrogel for the effective antigen delivery. Intranasal immunization with ghrelin-PspA vaccine elicited serum immunoglobulin G antibodies against ghrelin and attenuated body weight gain in diet-induced obesity mice. This obesity-attenuating effect was caused by a decrease in fat accumulation and an increase in energy expenditure that was partially due to an increase in the expression of mitochondrial uncoupling protein 1 in brown adipose tissue. The development of this nasal vaccine provides a new strategy for the prevention and treatment of obesity.

Critical role of TSLP-responsive mucosal dendritic cells in the induction of nasal antigen-specific IgA response.

Thymic stromal lymphopoietin (TSLP) is an interleukin-7 (IL-7)-like cytokine involved in T helper 2 type immune responses. The primary target of TSLP is myeloid dendritic cells (DCs), however, little is known about the mechanism by which TSLP elicits respiratory IgA immune responses upon mucosal immunization. Here, we found that the levels of TSLP and TSLPR were upregulated in the mucosal DCs of mice nasally immunized with pneumococcal surface protein A (PspA) plus cholera toxin (CT) compared with those immunized with PspA alone. PspA-specific IgA responses, but not IgG Ab responses were significantly reduced in both serum and mucosal secretions of TSLPR knockout mice compared with wild-type mice after nasal immunization with PspA plus CT. Furthermore, CD11c+ mucosal DCs isolated from TSLPR knockout mice nasally immunized with PspA plus CT were less activated and exhibited markedly reduced expression of IgA-enhancing cytokines (e.g., APRIL, BAFF, and IL-6) compared with those from equivalently immunized wild-type mice. Finally, exogenous TSLP promoted production of IgAs in an in vitro DC-B cell co-culture system as exhibited by enhanced IL-6 production. These results suggest that TSLP-TSLPR signaling is pivotal in the induction of nasal respiratory immunity against pathogenic pneumococcal infection.

The hnRNP-Htt axis regulates necrotic cell death induced by transcriptional repression through impaired RNA splicing.

In this study, we identify signaling network of necrotic cell death induced by transcriptional repression (TRIAD) by α-amanitin (AMA), the selective RNA polymerase II inhibitor, as a model of neurodegenerative cell death. We performed genetic screen of a knockdown (KD) fly library by measuring the ratio of transformation from pupa to larva (PL ratio) under TRIAD, and selected the cell death-promoting genes. Systems biology analysis of the positive genes mapped on protein-protein interaction databases predicted the signaling network of TRIAD and the core pathway including heterogeneous nuclear ribonucleoproteins (hnRNPs) and huntingtin (Htt). RNA sequencing revealed that AMA impaired transcription and RNA splicing of Htt, which is known as an endoplasmic reticulum (ER)-stabilizing molecule. The impairment in RNA splicing and PL ratio was rescued by overexpresion of hnRNP that had been also affected by transcriptional repression. Fly genetics with suppressor or expresser of Htt and hnRNP worsened or ameliorated the decreased PL ratio by AMA, respectively. Collectively, these results suggested involvement of RNA splicing and a regulatory role of the hnRNP-Htt axis in the process of the transcriptional repression-induced necrosis.

Nucleotide sequence of the genes encoding the L3, L4, and L23 equivalent ribosomal proteins from the archaebacterium Halobacterium halobium.

A lambda EMBL clone containing a gene cluster coding for the ribosomal proteins L3, L4, L23 and 5' region of L2 was identified in a genomic library for the halophilic archaebacterium Halobacterium halobium using a heterologous hybridization probe from the related organism Halobacterium marismortui. The clone also contains two conserved open reading frames found in H. marismortui, although with still unknown function. Its gene organization is very similar to that of 'S10 operon' of H. marismortui. The deduced amino acid sequence of these ribosomal proteins (HhaL3, HhaL4, HhaL23 and 5' region of HhaL2) shows high similarity (64-71%) to those of the archaebacterium H. marismortui and a lesser degree of similarity to their eukaryotic (31-42%) and eubacterial (17-33%) counterparts.

Charge isomers of urinary bikunin (trypsin inhibitor).

It was observed that the purified urinary bikunin (trypsin inhibitor) consisted of four major isomers with different electric charges which could be separated by HPLC using a Mono Q column. These isomers revealed the same antitrypsin activity and did not show any differences in the apparent molecular weight by SDS-PAGE, amino-acid composition, N-terminal amino-acid sequence (1-40) and C-terminal amino acid (Leu). The contents of sialic acid and uronic acid were also identical among these isomers. However, analysis of chondroitin sulfate revealed all the glycosaminoglycan chains of these isomers were undersulfated, comprising nonsulfated and 4-sulfated disaccharide units, and 4-sulfated disaccharide unit ratio varied among these isomers. After the chondroitin ABC lyase digestion, all the isomers were eluted at the same position on a Mono Q column chromatography. These results indicated that charge isomers of urinary bikunin was attributed to the difference on sulfation ratio in a glycosaminoglycan chain.

Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques.

We previously established a nanosized nasal vaccine delivery system by using a cationic cholesteryl group-bearing pullulan nanogel (cCHP nanogel), which is a universal protein-based antigen-delivery vehicle for adjuvant-free nasal vaccination. In the present study, we examined the central nervous system safety and efficacy of nasal vaccination with our developed cCHP nanogel containing pneumococcal surface protein A (PspA-nanogel) against pneumococcal infection in nonhuman primates. When [(18)F]-labeled PspA-nanogel was nasally administered to a rhesus macaque (Macaca mulatta), longer-term retention of PspA was noted in the nasal cavity when compared with administration of PspA alone. Of importance, no deposition of [(18)F]-PspA was seen in the olfactory bulbs or brain. Nasal PspA-nanogel vaccination effectively induced PspA-specific serum IgG with protective activity and mucosal secretory IgA (SIgA) Ab responses in cynomolgus macaques (Macaca fascicularis). Nasal PspA-nanogel-induced immune responses were mediated through T-helper (Th) 2 and Th17 cytokine responses concomitantly with marked increases in the levels of miR-181a and miR-326 in the serum and respiratory tract tissues, respectively, of the macaques. These results demonstrate that nasal PspA-nanogel vaccination is a safe and effective strategy for the development of a nasal vaccine for the prevention of pneumonia in humans.

Fragmentation of the beta-subunit of human chorionic gonadotropin produced by choriocarcinoma.

When human chorionic gonadotropin (hCG) was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions with dithiothreitol (DTT), a smaller weight material (CTP'), in addition to the beta-subunit, could be detected by Western blot analysis using antiserum for hCG beta-carboxy-terminal peptide (CTP). The CTP' band was much more apparent with urinary hCG from a patient with choriocarcinoma than with that from normal pregnant women. Second-dimensional electrophoresis of the choriocarcinoma hCG (c-hCG) after reduction with DTT indicated that the CTP', Mr 25,000, was released from the beta-subunit. The carbohydrate structure of the CTP' was analyzed by affinity with lectin-peroxidase on a nitrocellulose membrane. The CTP' did not interact with Concanavalin A, but exhibited strong interaction with both RCA120 and Arachis hypogaea after removal of sialic acid, indicating that it was released as a fragment containing an O-linked sugar chain as was found in the hCG beta carboxy-terminal portion. Western blot analysis using the antisera for hCG alpha, hCG beta, and hCG beta-CTP showed that the CTP' contains not only the carboxy-terminal portion but also a part of the internal (core) portion of the beta-subunit molecule. This dissociation of the c-hCG beta was further supported by the presence of a faster moving component (FMC) which may correspond to the NH2-terminal side counterpart. The desialylated FMC could be detected by Concanavalin A and RCA120 but not by Arachis hypogaea, indicating that it contains N-linked rather than O-linked sugar chains. The FMC does not contain any of the epitopes for the antisera examined in Western blot. These results indicate that the beta-subunit of the choriocarcinoma urine hCG has an unusual site which is dissociated into two components of Mr 25,000 (CTP') and Mr 18,000 (FMC) by DTT reduction.

ELISA for a complexed antigen with a monoclonal antibody blocking reaction with the free antigen-assay-specific for complexed prostate-specific antigen.

We have developed an enzyme-linked immunoassay (ELISA) for serum complexed-antigen (prostate-specific antigen; PSA)(c-PSA) with simultaneous blocking of free-antigen (PSA)(f-PSA). The assay utilizes three different monoclonal antibodies (MAbs) recognising three distinct PSA epitopes. The detection limit was established as 0.19 microg/l (n=20, mean of zero standard+2S.D.) and the average recovery of f-PSA was 98-100%. The within-run and between-day coefficients of variation (CV) ranged from 2.1% to 3.2% and 2.8% to 6.3%, respectively. There was a good correlation between serum c-PSA measured by the present ELISA and PSA-alpha(1)-antichymotrypsin complex (PSA-ACT) concentrations (r=0.991). This method should provide a better tool for discriminating between benign and malignant prostatic disease.

Surgery of giant bulla with tube drainage and bronchofiberoptic bronchial occlusion.

A case of emphysematous bullae and right lung cancer is presented. At first, left giant bulla was managed by minithoracotomy and tube drainage combined with bronchofiberoptic bronchial occlusion to preserve the respiratory function. Four weeks later, right lung cancer was successfully resected.

Right anterior caudocranial oblique projection for portal venography; its indications and advantages.

Right anterior caudocranial oblique (RACCO) projections were evaluated for transarterial portography with digital subtraction angiography. For RACCO projection, the image intensifier was tilted 25 degrees caudally and 30 degrees to the patient's right with the patient in the supine position. The abnormal findings imaged in 20 patients were analyzed by types of abnormalities and their locations in the portal venous systems by comparing RACCO and PA projections. The RACCO view was superior to the PA projection for demonstrating either encasements or tumor thrombi in relatively proximal segments of the portal venous branches. Two encasement lesions and a tumor thrombus were imaged only on the RACCO projection. However, the RACCO projection had no definite advantage over the PA view for showing encasements or tumor thrombi in the distal segmental branches. The RACCO projection was superior to the PA view for demonstrating defects in the hepatograms of some hepatic segments (S5 and S8). We concluded that the RACCO view is extremely useful and is indicated for angiographic work-ups of hepatic or biliary lesions when abnormalities are suspected in proximal portal venous branches.

Early and delayed imaging with 123I-IMP SPECT in patients with ischemic cerebrovascular disease.

Cerebral blood flow imaging with N-isopropyl (I-123) p-iodoamphetamine (IMP) was performed in 44 patients with ischemic cerebrovascular disease (CVD) at an acute or subacute stage less than 30 days from the onset. IMP imaging was obtained at 20 minutes (early scan) and 4 hours (delayed scan) after intravenous injection of 222 MBq of IMP. The region of interest (ROI) was selected in a slice compatible with the findings on the CT images, and the lesion to tissue ratio (L/T ratio) was calculated in a comparison with the unaffected side. The redistribution index (RI) was also calculated by dividing the difference between the L/T ratio in early and delayed image by the L/T ratio in the early image. The patients were classified into three groups (Grade 1, 2, 3) on the basis of the CT findings. The L/T ratio in the delayed images and RI was high in grade 1 and 2 groups and low in grade 3 groups both in early and delayed scans. The RI had tendency to grow high as the days after the onset became later. In the duration period from 4 to 7 days, 'reversed' redistribution was observed in 4 cases. Follow up examinations were performed in 6 cases in grade 3 group. The RI became higher in 3 cases and lower in 3 cases in the second examination. In conclusion, good redistribution was observed in grade 1 and 2 groups, and the prognosis was good. On the other hand, poor redistribution was observed in grade 3 group.(ABSTRACT TRUNCATED AT 250 WORDS)

Retroperitoneal leiomyosarcoma. Its MR manifestations.

The magnetic resonance manifestations of retroperitoneal leiomyosarcomas in two patients are described. The tumors were isointense to muscle on T1-weighted images and exhibited a relatively high signal on T2-weighted images. Postcontrast T1-weighted images after the administration of gadolinium-dietilenetriaminepentaacetic acid produced moderate enhancement except in regions of central necrosis.

Pseudo lesions on CTAP secondary to arterio-portal shunts.

PURPOSE: To clarify the characteristics of the defects on computed tomography (CT) during arterial portography (CTAP) due to arterio-portal shunting (APS). METHOD AND PATIENTS: The defects in the hepatic parenchyma on CTAP due to angiography proven APS were characterized according to their configuration, size, sharpness of margins, location of the lesion in the liver, and homogeneity of the lesions. RESULTS: Twenty five patients had non-tumor related APS. The shunts were single on angiography in 15 cases and multiple in 10 cases. The APS were due to liver cirrhosis in 22 patients, and to trauma in the other three. The lesions were irregular in shape in nine cases, wedge shaped in six cases, and round in four cases. In three cases both wedge and round lesions were seen. The margins were sharp in five single defects in five cases. The rest of the lesions were judged to have unsharp margins except for one case in which both clear and unclear lesions were present. In 10 cases, internal enhancement of the lesions were seen on CTAP. The location of the defects were central in five cases, peripheral in nine, and both peripheral and central in eight cases. CONCLUSION: The parenchymal defects due to APS were typically irregular in shape, with unsharp margins and internal enhancement.

Repositioning of a displaced catheter for percutaneous hepatic arterial infusion chemotherapy.

This is a report of a patient with hepatocellular carcinoma whose hepatic arterial catheter, which was introduced via a subclavian arterial branch, migrated into his right gastric artery. The catheter was successfully maneuvered to a suitable position without any further invasive procedure. The importance of preparatory embolization of the right gastric artery is underscored.

Computed tomography for the assessment of blunt abdominal trauma.

Computed tomography (CT) was carried out on 70 patients with blunt abdominal trauma. Damage to the abdominal organs was clearly demonstrated by CT. The positive rates of CT in traumatic lesions were higher than those of ultrasound (US). Intraperitoneal hemorrhage was well demonstrated by US. Serum GPT levels were significantly elevated in patients with liver damage (p less than 0.001). CT proved to be useful for detecting damage to organs in patients with abdominal trauma.

Percutaneous transluminal renal angioplasty of arterial divarication: application of kissing balloon technique.

We present a patient with renal artery stenoses due to fibromuscular dysplasia involving the point of arterial bifurcation that we could successfully dilate with percutaneous transluminal angioplasty. We used the kissing balloon technique to prevent possible complications of percutaneous transluminal renal angioplasty (PTRA), including occlusion of the other branch while dilating one branch. The kissing balloon technique worked well for PTRA of the arterial bifurcation.

Lipiodolized gelatin sponge: a simple method to make gelatin sponge radiopaque.

Gelatin sponge particles were rendered radiopaque by soaking them in small amounts of iodized oil (LP-GS). They were evaluated for their usefulness in transarterial embolization (TAE). The LP-GS particles were well visualized fluoroscopically during TAE. The duration of the opacity of LP-GS exceeded 24 hours both in vitro and in vivo. LP-GS are useful, and are an easily obtainable, radiopaque, embolic material.

Pseudoaneurysm of the celiac artery demonstrated by radionuclide angiography.

The usefulness of radionuclide angiography in patients with vascular occlusion or aneurysm is well documented. In this paper, a rare case of pseudoaneurysm of the celiac artery demonstrated by radionuclide angiography is reported.