Humoral immune response associated with lyme borreliosis in nonhuman primates: analysis by immunoblotting and enzyme-linked immunosorbent assay with sonicates or recombinant proteins

The immune response to Borrelia burgdorferi, the causative agent of Lyme disease, is complex. We studied the immunoglobulin M (IgM) and IgG antibody response to N40Br, a sensu stricto strain, in the rhesus macaque(nonhuman primate [NHP]) model of infection to identify the spirochetal protein targets of specific antibody. Antigens used in enzyme-linked immunosorbent assays were whole-cell sonicates of the spirochete and recombinant proteins of B. burgdorferi. Immunoblotting with a commercially available strip and subsequent quantitative densitometry of the bands were also used. Sera from four different groups of NHPs were used: immunocompetent, transiently immunosuppressed, extended immunosuppressed, and uninfected. In immunocompetent and transiently immunosuppressed NHPs, there was a strong IgM and IgG response. Major proteins for the early IgM response were P39 and P41 and recombinant BmpA and OspC. Major proteins for the later IgG response were P39, P41, P18, P60, P66, and recombinant BmpA and DbpA. There was no significant response in the NHPs to recombinant OspA or to Arp, a 37-kDa protein that elicits an antibody response during infection in mice. Most antibody responses, except for that to DbpA, were markedly diminished by prolonged dexamethasone treatment. This study supports the hypothesis that recombinant proteins may provide a useful adjunct to current diagnostic testing for Lyme borreliosis.     

Antibody responses in patients with staphylococcal septicemia against two Staphylococcus aureus fibrinogen binding proteins: clumping factor and an extracellular fibrinogen binding protein

We analyzed the serum antibody responses against two Staphylococcus aureus fibrinogen binding proteins, the cell-bound clumping factor (Clf) and an extracellular fibrinogen binding protein (Efb). The material consisted of 105 consecutive serum samples from 41 patients suffering from S. aureus septicemia and 72 serum samples from healthy individuals. An enzyme-linked immunosorbent assay (ELISA) was developed. Healthy individuals showed variable levels of antibodies against the studied antigens, and cutoff levels (upper 95th percentile) against these antigens were determined. No correlation was seen between serum antibody levels against Clf and Efb. In acute-phase samples 27% of patients showed positive antibody levels against Clf and 10% showed positive levels against Efb, while in convalescent-phase samples 63% (26 of 41) showed a positive serology against Clf and 49% (20 of 41) showed a positive serology against Efb. Antibody levels against Efb were significantly lower in the acute-phase sera than in sera from healthy individuals (P = 0. 002). An antibody response against Clf was most frequent in patients suffering from osteitis plus septic arthritis and from endocarditis (80% positive). The antibody response against Efb appeared to develop later in the course of disease. A possible biological effect of measured antibodies was demonstrated with the help of an inhibition ELISA, in which both high-titer and low-titer sera inhibited the binding of bacteria to fibrinogen. In conclusion, we have demonstrated in vivo production of S. aureus fibrinogen binding proteins during deep S. aureus infections and a possible diagnostic and prophylactic role of the corresponding serum antibodies in such infections.     

Nine novel DNA components associated with the foorkey disease of large cardamom: Evidence of a distinct babuvirus species in Nanoviridae

Foorkey disease is a serious constraint to the production of large cardamom (Amomum subulatum, family Zingiberaceae). The disease is characterized by profuse proliferation of excessive stunted shoots, which makes the clump totally unproductive. The disease has been known in India since 1936 but the complete genome of the virus had not yet been characterized. In a preliminary study, an associated virus tentatively named as Cardamom bushy dwarf virus (CBDV) was identified based on the partial sequence of a single DNA component (DNA-R). In the present study, a high incidence (37.2–39.3%) of foorkey was recorded in certain plantations in the Darjeeling hills located at lower altitudes (300–1380 m) and CBDV was detected in several field samples by PCR. Nine novel DNA components were isolated and characterized from foorkey affected plants. CBDV contained six major DNA components (DNA-R, -S, -M, -C, -N and -U3) similar to the integral genome components known for the members of the genus Babuvirus in the family Nanoviridae. Additional components, satellite Rep (DNA-sRep1) and unknown components (DNA-Uf1 and -Uf2) were also identified. The size of the genome components ranged from 1028 to 1127. The sequence identity and phylogeny based on the individual components as well as overall genome (59.8–62% identity) distinguished CBDV from the two existing babuvirus species, Banana bunchy top virus and Abaca bunchy top virus. CBDV is the first distinct babuvirus species that affects plant species outside family Musaceae. This study shows further diversity in the genus Babuvirus.     

Traveler''s diarrhea associated with rotavirus infection: analysis of virus-specific immunoglobulin classes.

An enzyme-linked immunosorbent assay for the detection of rotavirus-specific immunoglobulin G (IgG), IgM, and IgA antibodies was used in a serological study of Traveler's diarrhea. The antigenically related simian rotavirus (SA-11) was used as antigen in this study. Serum was obtained from two groups of volunteers before travel (pre-travel) and at various times after arrival at their destination (post-travel). One group consisted of 47 American Peace Corps volunteers who traveled to Honduras, and the second group consisted of 66 Panamanian travelers who visited Mexico. An association between rotavirus and Traveler's diarrhea was found in each group with 36% of American Peace Corps volunteers and 30% of Panamanians with diarrhea demonstrating a greater than or equal to 4-fold increase in rotavirus antibody titers in the post- as compared to the pre-travel specimens. While no rotavirus-specific IgM antibody was detected in any serum tested, increases in specific antibody were found in both the IgG and IgA immunoglobulin classes.     

Antigenicity of mouse interferons: distinct antigenicity of the two L cell interferon species

Antibodies were raised in rabbits against Newcastle disease virus-induced L cell interferon of high purity, and against each of its two major species, F(24K) and S(36K) interferons. The two interferons were found antigenically distinct. Thus, the anti-F and anti-S sera failed to neutralize appreciably the antiviral, as well as the cell growth-inhibiting, activity of the heterologous interferon. Heterologous reactions were also undetectable in a modified neutralization test, in which secondary antibody against rabbit γ-globulin was used to remove, before the assay for the residual interferon, any interferon-antibody complexes that might remain biologically active. Affinity chromatography of interferons on immobilized antibodies also showed antigenic distinctness of F and S interferons. Poly (I) ·poly (C)-induced L cell interferon and Newcastle disease virus-induced C₂₄₃ cell interferon also consisted of two distinct species which are antigenically similar to F and S.     

Genetic evidence for the presence of two distinct hantaviruses associated with Apodemus mice in Croatia and analysis of local strains

In Europe, Dobrava-Belgrade (DOBV), Saaremaa (SAAV), and Puumala (PUUV) viruses are known to cause hemorrhagic fever with renal syndrome (HFRS). All three hantaviruses are now found in Croatia. Lung tissue samples of 315 Apodemus mice trapped in 2003-2004 were screened for the presence of hantaviral N-Ag and 20 mice (6.3%) were found either strongly positive or weak/suspected-positive. Partial sequences of hantavirus M and S segments were recovered by RT-PCR from six mice and subjected to (phylo)genetic analysis that revealed the presence of four novel strains of DOBV and one of SAAV. Curiously, one of the newly described DOBV strains was found in Apodemus agrarius mouse, that is, not in the traditional host, A. flavicollis mice, suggesting a spillover event. S segment sequences recovered previously from HFRS cases [Markoti? et al., 2002] were confirmed as DOBV sequences; one of which appeared particularly close to the prototype Slovenian DOBV isolate. Taken together with earlier data on PUUV in Croatia, these results show a co-circulation of three European hantavirus pathogens in this country. So far, not a single SAAV sequence has been recovered from HFRS patients either in Croatia or neighboring Slovenia and Hungary nor in Slovakia suggesting a somewhat lower fequency of acute SAAV infection in humans in this part of Europe than for example in the Baltics.     

Endotoxic-lipopolysaccharide-specific binding proteins on lymphoid cells of various animal species: association with endotoxin susceptibility.

Endotoxic lipopolysaccharide (LPS), a common structural component of all gram-negative bacteria, is well recognized for its capacity to interact with and perturb immunologically relevant cells. Using a radioiodinated, photoactivatable LPS probe, we have recently identified an 80-kilodalton LPS-specific binding protein on murine B lymphocytes. We now have extended these studies to determine if other mammalian species, as well as representative endotoxin-resistant species (frog and chicken), have a similar LPS-binding protein. We have identified what appears to be a relatively conserved 80-kilodalton LPS-binding protein on mononuclear cells of all mammalian species tested. However, both frog and chicken leukocytes failed to show the presence of a similar LPS-binding protein. It is possible that the presence of specific LPS-binding proteins may be important for endotoxin sensitivity of most mammalian species.     

Sequence analysis of RNA-2 of different isolates of broad bean wilt virus confirms the existence of two distinct species

Summary.  The complete nucleotide sequence of RNA-2 from a Japanese isolate IP of broad bean wilt virus (BBWV) was determined. The sequence encodes a single large polyprotein, which contains a putative movement protein and two coat proteins (CPs). The 3′-terminal sequences of RNA-2 were also determined for three other Japanese isolates and two ATCC isolates (PV132 and PV176) of BBWV. The CPs of the four Japanese isolates share 86.8–98.0% amino acid sequences homology with one another and 88.3–96.5% with those reported for the isolate PV131 (BBWV-2). However, they have only 57.9–66.2% homology with those of PV132 and PV176 (BBWV-1). Received December 7, 1998 Accepted February 23, 1999     

Demonstration of factor H-like protein 1 binding to Treponema denticola, a pathogen associated with periodontal disease in humans

Treponema denticola is an important contributor to periodontal disease. In this study we investigated the ability of T. denticola to bind the complement regulatory proteins factor H and factor H-like protein 1 (FHL-1). The binding of these proteins has been demonstrated to facilitate evasion of the alternative complement cascade and/or to play a role in adherence and invasion. Here we demonstrate that T. denticola specifically binds FHL-1 via a 14-kDa, surface-exposed protein that we designated FhbB. Consistent with its FHL-1 binding specificity, FhbB binds only to factor H recombinant fragments spanning short consensus repeats (SCRs) 1 to 7 (H7 construct) and not to SCR constructs spanning SCRs 8 to 15 and 16 to 20. Binding of H7 to FhbB was inhibited by heparin. The specific involvement of SCR 7 in the interaction was demonstrated using an H7 mutant (H7AB) in which specific charged residues in SCR 7 were replaced by alanine. This construct lost FhbB binding ability. Analyses of the ability of FHL-1 bound to the surface of T. denticola to serve as a cofactor for factor I-mediated cleavage of C3b revealed that C3b is cleaved in an FHL-1/factor I-independent manner, perhaps by an unidentified protease. Based on the data presented here, we hypothesize that the primary function of FHL-1 binding by T. denticola might be to facilitate adherence to FHL-1 present on anchorage-dependent cells and in the extracellular matrix.     

Comparative analysis of serum proteomes: discovery of proteins associated with osteonecrosis of the femoral head

No means exist to evaluate the activity status, turnover, and prognosis of idiopathic osteonecrosis of the femoral head (IONFH) except for X-ray evidence of segmental collapse as a very good marker for prognosis. Moreover, the only current method for diagnosis of this disease is through physical examination and diagnostic imaging results, and no serum biochemical markers exist. A comparative analysis of serum proteomes was performed to discover proteins associated with osteonecrosis of the femoral head. Two-dimensional electrophoresis (2-DE) patterns of human sera from 10 patients with IONFH and 10 normal subjects were analyzed. The differentially expressed spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and 7 proteins were found. The expression levels of tissue-type plasminogen activator (t-PA), bone-carboxyglutamate protein (BGP), c-sis, and an unknown protein were downregulated in the sera of patients with IONFH, whereas the other 3 proteins, including plasminogen activator inhibitor type 1 (PAI-1), crosslaps, and anti-p53 antibody, were upregulated. To examine their applicability as diagnostic markers, levels of the 6 identified proteins in serum were validated from patients with IONFH, osteoarthritis (OA), rheumatoid arthritis (RA), and fracture using the enzyme-linked immunosorbent assay (ELISA) method. It was found that only serum levels of t-PA, PAI-1, crosslaps, and anti-p53 antibody in patients with IONFH were always significantly different from those in patients with OA, RA, and fracture. These results suggest that serum levels of t-PA, PAI-1, crosslaps, and anti-p53 antibody could be used as noninvasive diagnostic biomarkers for IONFH.     

Endometriosis: Insulin-like growth factor binding proteins in peritoneal fluid of women with minimal and mild endometriosis

This prospective cohort study was carried out in a university-basedInfertility clinic to determine the profile of insulin-likegrowth factor binding proteins (IGFBPs) in patients with mildendometriosis and no obvious mechan ical factor contributingto infertility. A total of 26 patients with minimal and mildendometriosis and 10 controls contributed peritoneal fluid atsurgery. The variety, expression and levels of IGFBPs were determinedby radio inununoassay and Western ligand blots (WLBs) with quantitationby laser densitometer. A 27 kDa species was significantly lowerand a 31 kDa species tended to be lower in patients with endometriosisas determined by quantitative laser densitometer. The levelsof IGFBP-3 detected by radioimmunoassay and by WLB were correlatedin the control group and in the patients with endometriosisin the follicular phase but not in patients with endometriosisin the luteal phase. The level of 27 kDa species seen on WLBsdid not appear to correspond to IGFBP-1 determined by radiomimunoassayand IGFBP-3 levels in luteal phase endometriosis patients alsodeparted from values determined by radioimmunoassay. These discrepancies suggest a complex system to control levels of IGFin the peritoneum involving multiple binding proteins and proteases.The IGFBPs of patients with endometriosis may contribute toreproductive dysfunction and be able to serve as markers.     

白塞病伴发恶性肿瘤41例临床分析

 目的 研究白塞病（BD）伴发恶性肿瘤的临床特点和相关性。方法 分析北京协和医院收治的BD伴发恶性肿瘤患者的临床资料。结果 1)BD伴发恶性肿瘤41例 (男16例,女25例)。其中恶性血液病29例,骨髓增生异常综合征占20例,白血病7例,另再生障碍性贫血2例和淋巴瘤1例;实体肿瘤13例,结直肠癌占3例（其中1例合并子宫内膜癌）,膀胱癌2例,食管癌、胃癌、胰腺癌、甲状腺癌、乳腺癌、宫颈癌、肾癌和转移性腺癌各1例。2)BD伴有恶性肿瘤的患者年龄偏大、消化道受累多见。3)BD伴恶性血液病和实体肿瘤两组间年龄、BD与肿瘤的间期以及肿瘤进展时BD的活动有显著差异。结论 BD伴发恶性肿瘤有一定的风险和特点,高龄、消化道受累可能是BD伴发恶性肿瘤的危险因素。二者的关联性尚不清楚,需要在临床工作进一步总结。     

Comprehensive analysis of the LRRK2 gene in sixty families with Parkinson's disease

Mutations in the gene leucine-rich repeat kinase 2 (LRRK2) have been recently identified in families with Parkinson's disease (PD). However, the prevalence and nature of LRRK2 mutations, the polymorphism content of the gene, and the associated phenotypes remain poorly understood. We performed a comprehensive study of this gene in a large sample of families with Parkinson's disease compatible with autosomal dominant inheritance (ADPD). The full-length open reading frame and splice sites of the LRRK2 gene (51 exons) were studied by genomic sequencing in 60 probands with ADPD (83% Italian). Pathogenic mutations were identified in six probands (10%): the heterozygous p.G2019S mutation in four (6.6%), and the heterozygous p.R1441C mutation in two (3.4%) probands. A further proband carried the heterozygous p.I1371 V mutation, for which a pathogenic role could not be established with certainty. In total, 13 novel disease-unrelated variants and three intronic changes of uncertain significance were also characterized. The phenotype associated with LRRK2 pathogenic mutations is the one of typical PD, but with a broad range of onset ages (mean 55.2, range 38-68 years) and, in some cases, slow disease progression. On the basis of the comprehensive study in a large sample, we conclude that pathogenic LRRK2 mutations are frequent in ADPD, and they cluster in the C-terminal half of the encoded protein. These data have implications both for understanding the molecular mechanisms of PD, and for directing the genetic screening in clinical practice.     

Intraductal papillary neoplasms of the bile duct consist of two distinct types specifically associated with clinicopathological features and molecular phenotypes

Intraductal papillary neoplasm of the bile duct (IPNB) is a grossly visible papillary biliary neoplasm with morphological variations and occasional invasion. Recently a new classification of IPNB into type 1 and type 2 was proposed in which the type 1 IPNBs consist of fine papillary neoplastic glands and the type 2 IPNBs consist of complex branching glands, seldom with foci of solid-tubular components. However, clinicopathological and molecular characteristics of these types of IPNBs are yet to be identified. We aimed to uncover clinicopathological and molecular characteristics of the types of IPNBs. Thirty-six IPNBs were studied retrospectively. Clinicopathological features as well as molecular alterations of 31 genes were evaluated by means of targeted next-generation sequencing and immunohistochemical examination of expression of mucin and cancer-associated molecules. The 36 IPNBs were classified into 22 of type 1 and 14 of type 2. The type 1 IPNBs were associated with a non-invasive phenotype, intestinal and oncocytic subtypes, development in the intrahepatic bile duct, overt mucin production, and a relatively good prognosis. The type 2 IPNBs were associated with an invasive phenotype, the pancreatobiliary subtype, development within the extrahepatic bile duct, and worse prognosis compared with the type 1 IPNBs. In the molecular analysis, recurrent mutations were found in TP53 (34.3%), KRAS (31.4%), STK11 (25.7%), CTNNB1 (17.1%), APC (14.3%), SMAD4 (14.3%), GNAS (11.4%), PBRM1 (11.4%), ELF3 (8.6%), KMT2C (8.6%), NF1 (8.6%), PIK3CA (8.6%), ARID1A (5.7%), ARID2 (5.7%), BAP1 (5.7%), BRAF (5.7%), EPHA6 (5.7%), ERBB2 (5.7%), ERBB3 (5.7%), KMT2D (5.7%), and RNF43 (5.7%). Mutations in KRAS and GNAS were enriched in the type 1 IPNBs, whereas mutations in TP53, SMAD4, and KMT2C were enriched in the type 2 IPNBs. These results indicate that IPNBs consist of two distinct types of neoplasms specifically associated with clinicopathological features and molecular phenotypes. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Phylogeny of capsid proteins of rod-shaped and filamentous RNA plant viruses: two families with distinct patterns of sequence and probably structure conservation

Computer-assisted comparative analysis of all available amino acid sequences of the capsid proteins of positive strand RNA plant viruses with helical capsids is described. Two distinct families of homologous proteins were delineated through statistically significant sequence similarities, one including the capsid proteins of rod-shaped viruses (tobamo-, tobra-, hordei-, and furoviruses) and the other those of filamentous viruses (poty-, bymo-, potex-, carla-, and closteroviruses). It was concluded that the capsid proteins of all rod-shaped viruses, on the one hand, and filamentous viruses, on the other hand, evolved from common ancestors. Analysis of residue conservation patterns in the capsid proteins of rod-shaped viruses revealed maintenance of the hydrophobic core and of the (putative) salt bridge between conserved Arg and Asp residues. Sequence comparisons within the filamentous virus family expanded the observations on the relationship between the capsid proteins of potex-, carla-, poty-, and bymoviruses. Grouping of the beet yellows closterovirus capsid protein sequence, recently determined in this laboratory (Agranovsky et al., J. Gen. Virol., 1991, 72, 15-23), with those of potex- and carlaviruses was demonstrated. The coat protein of another closterovirus, apple chlorotic leaf spot virus, appeared to constitute a distinct phylogenetic lineage. Despite the lack of significant overall similarity, comparison of the alignments of the capsid proteins of the two families suggested formation of analogous salt bridges.     

Inflammatory pseudotumor of the liver associated with malignant disease: report of two cases and a review of the literature

Two cases of inflammatory pseudotumor (IPT) of the liver associated with gastrointestinal tract cancer are reported. In addition, the etiological correlation between IPT and abscesses of the liver in cancer patients is discussed. The first patient was a 63-year-old woman who underwent distal gastrectomy and partial hepatectomy under a diagnosis of stomach cancer with liver metastasis. The second patient was a 66-year-old man who had undergone surgery for rectal cancer 6 years previously and underwent partial hepatectomy under a diagnosis of metastasis of rectal cancer to the liver. The gastric cancer was a papillary adenocarcinoma limited to the mucosa, and the rectal cancer was a moderately differentiated adenocarcinoma limited to the subserosa. The resected liver tumor in the first case measured 5.5×5.0×4.0 cm and was 2.5×1.9×1.6 cm in the second. The cut surface showed that both masses were well circumscribed and divided into lobules by fibrous tissue. They were yellowish white in color and there was no evidence of necrosis or hemorrhage. Histologically the masses consisted of fibrous areas and cellular areas, and the cellular areas consisted of fascicles of plump spindle cells mingled with varying numbers of plasma cells, lymphocytes, and histiocytes. The masses were diagnosed as IPTs. Obliterating phlebitis suggesting infection via the portal vein was seen in the adjacent liver tissue in both cases. According to previous cases reported in the literature, there are three types of cancers associated with hepatic IPT: gastrointestinal tract cancer, biliary tract cancer, and cancers that need strong systemic chemotherapy. The underlying cancer types of IPT of the liver are almost similar to those associated with pyogenic liver abscesses suggesting the etiological correlation between IPT and abscesses of the liver.