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The objective of the study was to determine the correlation between group A streptococcal antigen detected from throat swabs with the culture results. A total of 1457 children had two swabs taken simultaneously, and culture and antigen detection were performed. There was a good correlation between antigen detection and isolation rates. In all, 225 strains of group A streptococcus were isolated;53 [57.6%] were from the 92 children with high antigen positivity, 68 [55.7%] were from the 122 children with medium antigen positivity and 77 [25.4%] were from 303 children with low antigen positivity;only 27 [2.9%] were from the 940 children with no antigen detected. We postulate that those who are antigen-positive, culture-negative carry the organisms in their throats, but they may be missed on culture because of the small number carried

We report on 2 patients who presented stiffness and spasms similar to those of stiff-man syndrome (SMS) that were limited to one leg for up to 11 years. Patients had serum glutamic acid decarboxylase (GAD) autoantibodies in high titer, clinical evidence of organ-specific autoimmunity, and electromyographic pattern of continuous motor unit activity with abnormally enhanced exteroceptive reflexes. The clinical and immunological profile suggests that this disorder may be a focal form of SMS.

Processing bodies (P-bodies) are cellular structures that have critical roles in mRNA degradation and post-transcriptional gene silencing. Some patients with autoimmune disease have high titer antibodies directed against P-bodies, and certain sera have been used as markers for the GW182 component of these structures. This study shows that available reference sera are unreliable markers for GW182 because the sera contain antibodies directed against Ge-1, a second P-body autoantigen.

We have developed a viral RNA (vRNA) dot blot assay for rapid identification of high-titer retrovirus vector production by packaging cell clones. The procedure employs Trizol LS reagent to purify vRNA from packaging cell supernatants, a sensitive dot blot assay, and Phosphorlmager technology to quantify packaged viral genomes in 2 days. Experiments performed on viral supernatants of known biological titer demonstrated that the vRNA dot blot assay was extremely sensitive and that dot intensity correlated directly with viral titer. It is often necessary to analyze approximately 100 virus producing cell clones, making this method useful as a rapid screen to identify the highest virus producing clones. The vRNA dot blot assay consistently identified a subset of candidate high-titer producer cell clones. In three independent screens the supernatant with the highest biological titer was produced by one of the previously defined candidate high-titer producer clones. Our procedure greatly facilitates virus titration by: (1) rapidly eliminating the vast majority of low-titer producer cell clones; (2) accurately identifying the subset of candidate high-titer producer clones for further biological titration and assessment of the proviral genomic structure; and (3) reducing laborious tissue culture manipulations to a minimum. Furthermore, the reliance of this method on molecular detection makes it ideally suited for the isolation of high-titer clones lacking a drug selection marker.

It has been generally estimated that useful quantitative characters may be fixed at the second generation of gynogenesis. Individual variations of agglutinating antibody production following an intraperitoneal injection with formalin-killed cells of Vibrio anguillarum serotype A were campared between gynogenetic (retaining 2nd polar body) and normal diploid. As a result, the range of the variation in gynogenetic fish was significantly wider than that of normal diploid. Gynogenetic and normal diploid fish resistant to the disease were selected by artificial infection. The next generations were produced by gynogenesis from survivors of the infected fish. Survival rate in a challenge test and rate of individuals showing high titer in the next generation of gynogenetic group were higher than those of normal diploid or non-selected control groups. In the 2nd generation of gynogenetic diploid selected by experimental infection, similar results were obtained. These results suggest that the fish resistant to vibriosis can be obtained at the 2nd generation of gynogenesis.

We report a patient with hemophilia A and high-titer factor VIII inhibitor who developed compartment syndrome of his forearm following trauma. Emergency fasciotomy was performed. Initial hemostatic treatment with factor VIII inhibitor bypassing activity (FEIBA) was unsuccessful. Bleeding was controlled with recombinant factor VIIa.

Chen Chu1,2, Aijie Xin1,3,4, Yuchuan Zhou1, and Yonglian Zhang1,3* Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China Graduate University of the Chinese Academy of Sciences, Shanghai 200031, China Shanghai Institute of Planned Parenthood Research, Shanghai 200031, China State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China *Correspondence address. Tel: þ86-21-54921163; Fax: þ86-21-54921011; E-mail: ylzhang@sibcb.ac.cn

Background Osteosarcomas are the most common primary bone malignancies found in children and adolescents. An optimized system was developed for efficient retroviral gene delivery into solid 143B osteosarcoma tumors in mice using a retargeted Env. In these studies, the viral Env CP was isolated from an in vitro screen of a library of feline leukemia virus Env randomized in the receptor-binding domain and maintained high titer on human 143B osteosarcoma cell line. Findings The vector developed to express the random Env libraries encoded the drug selectable marker neo. To adapt this for studies in live animals, the murine based vector was modified to express the luciferase gene. The bicistronic vector developed expressed both the CP Env and luciferase in the presence of either the MPMV CTE or a WPRE element. Virus bearing the CP FeLV Env variant maintained high titers after concentration allowing for direct visualization of delivery of the luciferase gene in subcutaneous 143B osteosarcoma tumors. Conclusion This system serves as a proof-of-concept for the use of novel FeLV Env pseudotyped MLV particles for in vivo gene delivery. Gene delivery and expression of lucerifase from viral particles bearing the CP Env was readily detected in live mice after a single round of intratumor injection.

We studied physical development, behavioral characteristics, and learning capacity in the offspring of mice immunized with nerve growth factor and bovine serum albumin. High titer of antibodies to these factors in the blood of pregnant females determines high levels of these antibodies in the blood of their pups. These changes modulate physical development, behavior, and learning capacity of rat pups. The effects of these antibodies differed in the strength and directionality. Antibodies to nerve growth factor more markedly retarded physical development, reduced learning capacity, and considerably increased pain thresholds in animals.

Background Baseline RF or ACPA positivity is reported to be linked with clinical response of some non-TNF bDMARDs in RA. However, its impact on the efficacy of TNF inhibitors was controversy. Objectives To evaluate the influence of baseline RF and ACPA “titers” on the patients characteristics and efficacy of infliximab (IFX) in RA patients using data of the RISING study, a randomized double-blind clinical study in MTX-refractory RA (ref-1, NCT00691028). Methods In the RISING study, after patients received 3 mg/kg IFX infusion at Weeks 0, 2, and 6, they were randomly assigned to infusion of 3, 6, or 10 mg/kg IFX, every 8 weeks from Week 14 to Week 46 in a double-blind manner (n=307, all patients received 3 mg/kg treatment until Week 14). Serum IFX levels and clinical responses based on DAS28-CRP were evaluated at Weeks 14 and 54. Results Both baseline RF and ACPA titers showed significant positive correlation with baseline plasma TNF level. We hypothesized that “both RF-High and ACPA-High” was correlated with higher TNF level. Then, we stratified the patients into 3 classes using baseline RF/ACPA titers; “Low/Low” (RF: Among above 3 classes, significant difference was observed in plasma baseline TNF level, which was previously reported to be closely correlated with low serum IFX level, and low clinical response (ref-2); low TNF level in Low/Low class, high TNF level in High/High class, and intermediate value in Middle class (median: 0.73, 1.15, and 0.91 pg/ml, respectively, p Conclusions High titers of both RF/ACPA at baseline is correlated with high baseline plasma TNF level which might be resulted in low IFX level and low clinical response in patients with continuing 3 mg/kg treatment, but not in patients receiving 6 or 10 mg/kg treatment. Baseline RF/ACPA might be the index of dose-escalation of IFX in RA. References Takeuchi T, et al. Mod Rheumatol 2009;19:478. ref-2: Takeuchi T, et al. Ann Rheum Dis 2011;70:1208. Disclosure of Interest T. Takeuchi Grant/research support from: AbbVie GK, Asahi Kasei Pharma, Astellas Pharma, AYUMI Pharmaceutical, Bristol-Myers KK, Chugai Pharmaceutical, Daiichi Sankyo, Eisai, Taisho Toyama Pharmaceutical, Takeda Pharmaceutical, Teijin Pharma, Consultant for: AbbVie GK, Asahi Kasei Medical, Astellas Pharma, AstraZeneca KK, Bristol-Myers KK, Daiichi Sankyo, Eli Lilly Japan, Janssen Pharmaceutical KK, Merck Serono, Mitsubishi Tanabe Pharma, Nippon Kayaku, Novartis Pharma KK, Takeda Pharmaceutical, Speakers bureau: AbbVie GK, Astellas Pharma, Bristol-Myers KK, Celtrion, Chugai Pharmaceutical, Daiichi Sankyo, Eisai, Janssen Pharmaceutical KK, Mitsubishi Tanabe Pharma, Nippon Kayaku, Pfizer Japan, Takeda Pharmaceutical, N. Miyasaka: None declared, T. Inui Employee of: Mitsubishi Tanabe Pharma, T. Yano Employee of: Mitsubishi Tanabe Pharma, T. Yoshinari Employee of: Mitsubishi Tanabe Pharma, T. Abe: None declared, T. Koike Consultant for: Eli Lilly Japan, Pfizer Japan, Speakers bureau: AbbVie GK, Astellas Pharma, Bristol-Myers KK, Chugai Pharmaceutical, Daiichi Sankyo, Eisai, Mitsubishi Tanabe Pharma, Pfizer Japan, Santen Pharmaceutical, Teijin Pharma, UCB Japan

The Japan Red Cross Blood Center tests Anti-HCV for screening in all of blood donors. The HCV-PHA Kit has been using for Anti-HCV screening since February 1991. The notification of HCV viremia is sent to the donor that showed anti-HCV high titer or s-GPT abnormal level. It is necesary to consult as the hepatitis C at an early stage and to prevent the another infection by blood transfusion.We examined MATRIX-HCV as Anti-HCV supplemental test with blood donor specimens. It was compared with the RIBA-II and HCV-RNA by RT-PCR method. The performance of MARTIX-HCV showed almost same as RIBA-II. Although total agreement between MATRIX-HCV and RIBA-II was 81.8% (63/77), the discrepant cases were near the cutoff value and seemed to be depended on the reactibity of NS3 and NS4 reagion. All 30 HCV-RNA positive cases out of 61 blood donors showed positive by MATRIX-HCV.

This article is available from http://www.labmedonline.org 2016, Laboratory Medicine Online This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Background: Throat culture is the golden standard for diagnosis of group A streptococcal (GAS) pharyngitis. However, because it is a time-consuming procedure, antibiotics are often empirically administrated. Rapid antigen tests (RATs) can detect bacterial infections within 15 minutes, thus helping to reduce unnecessary administration of antibiotics. Methods: In total, 108 patients, between 3 and 17 yr of age, who visited our hospital from August 2011 to July 2012, were tested for suspected acute pharyngitis with two RATs––SD Bioline Strep A (SD, Korea) and BinaxNOW Strep A (Binax, Inc., USA)––as well as throat culture. We compared the sensitivity, specificity, and consistency of the two RATs and assessed the clinical manifestations of GAS pharyngitis. Results: Of the 108 patients, 15 were confirmed to have GAS pharyngitis by throat culture. The SD test showed a sensitivity of 93.3% and a specificity of 97.8%; the positive and negative predictive values were 87.5% and 98.9%, respectively. The Binax test showed a sensitivity of 86.7% and a specificity of 100%; the positive and negative predictive values were 100% and 97.9%, respectively. The Kappa values for conformity degree were high, 0.887 and 0.918 in the SD and the Binax tests, respectively (P =0.00). Clinical manifestation assessment of GAS pharyngitis indicated that scarlatiniform rash and strawberry tongue were significantly associated signs (P

Background Immune tolerance induction (ITI) can reduce inhibitors against factor VIII concentrates by 70-80%. In this study, we elucidated the characteristics of inhibitors and attempted to determine the proper indications and timing for ITI. Methods Subjects included hemophilia A patients registered at the Korea Hemophilia Foundation from 1991 through 2014. Inhibitors were classified as persistent and transient. Patients were classified into groups according to peak inhibitor titer: low (

Author(s): Chaillon, A; Wack, T; Braibant, M; Mandelbrot, L; Blanche, S; Warszawski, J; Barin, F

Presentation of Case A 19-year-old man was admitted to the hospital on New Year's Day because of ascending paresthesias, progressive weakness of the legs, and a rash. He had been well until 10 days earlier, when a sore throat, productive cough, sweats, fever with a rise in the temperature to 39.4 °C, and dizziness developed. A physician found pharyngeal injection with questionable tonsillar exudates, and amoxicillin was prescribed. A rapid test for group A streptococcal antigen was negative. Five days before admission, the patient felt better, discontinued the amoxicillin, and returned to work. Three days later he began to have . . .

This case reports documents the progressive development of chronic interstitial cystitis during the overlapping process from rheumatoid arthritis to lupus. Concomitantly high titer antinuclear antibody with an anti-68 KD RNP pattern and other biological markers of lupus were observed in the blood. The symptoms dramatically improved under methotrexate therapy. The pathogenic mechanism is discussed.

Process development groups defining purification processes for therapeutic monoclonal antibodies are finding themselves at a crossroads. High titer feedstocks have challenged processes and facilities to recover single batches in excess of 50 kg, and triggered debates regarding the future direction of mAb purification. Are novel unit operations required to handle these high productivity processes in a cost-effective manner, or will the new generation of resins and membranes be sufficient? This workshop reviewed the state-of-the art in cGMP antibody purification processes, economics and cost of goods considerations, and facility fit issues surrounding novel as well as conventional purification technologies. In addition, pressures to adopt a mAb processing platform continue to mount, most recently through the introduction of QbD and an increasing frequency of multi-product facilities switching between legacy and newly launched mAbs. As modern processes become more industrialized (by convergence on a common set of highly-efficient, scalable, cost-effective unit operations across the industry), development strategies will need to be refined. If further advances in mAb bioprocessing are needed, where should companies focus their technology development efforts for downstream processing?

Protein engineering has for decades been a powerful tool in biotechnology for generating vast numbers of useful enzymes for industrial applications. Today, protein engineering has a crucial role in advancing the emerging field of synthetic biology, where metabolic engineering efforts alone are insufficient to maximize the full potential of synthetic biology. This article reviews the advancements in protein engineering techniques for improving biocatalytic properties to optimize engineered pathways in host systems, which are instrumental to achieve high titer production of target molecules. We also discuss the specific means by which protein engineering has improved metabolic engineering efforts and provide our assessment on its potential to continue to advance biology engineering as a whole.

Objective The aim of this study was to investigate the prevalence of autoantibodies in various liver diseases and evaluate the performance parameters of AMA, ANA and their combinations for PBC. Design and methods This study enrolled 115 PBC patients, 695 patients with other liver diseases and 105 healthy subjects. Results AMA were detected mainly in PBC patients, while ANA were detected in all categories. The high titer and multiple nuclear dot, rim-like and centromere (MRC) patterns of ANA were found in the majority of PBC patients, in contrast to moderate titers and other patterns. All the liver diseases but HAV and NAFLD had significantly independent association with ANA. The combination of MRC patterns and AMA showed superior performance parameters for PBC. Conclusions Although ANA can occur in various liver diseases, high titer and MRC patterns of ANA are more specific for PBC. The combination of MRC patterns and AMA has superior performance for PBC.

Heterophilic antibodies are still an important source of interference in immunoassays. We have conducted a screening study for interference in a panel of commercially available assays using two sera known to contain high titer Fc-reactive heterophilic antibodies.The sera were distributed to laboratories participating in the Nordic External Quality Assessment cooperation (EQANord). Duplicate samples pre-blocked with aggregated murine monoclonal MAK33 were also supplied. Discrepancies (>50%) between the results for native and blocked samples were used to classify the tested assays as susceptible to interference. A total of 170 different assay kits covering 91 analytes were tested.We found that 21 assays, covering 19 different analytes, were susceptible to interference from the heterophilic antibodies in the two sera. Many of these are clinically and commercially important assays. Some of the false results were grossly elevated and could have been detrimental to patient care in a clinical setting.Heterophilic antibodies with Fc-reactivity remain a threat. A more widespread use of antibody fragments and aggregated immunoglobulin could potentially improve the heterophilic antibody resistance of assays intended for clinical use.

During the current period the following key objectives were achieved: demonstration of high titer antibody production by geese following immunization with inactived H1N1 virus; completion of the epitope mapping of West Nile Virus-specific goose antibodies and initiation of epitope mapping of H1N1 flu-specific goose antibodies; advancement in scalable purification of goose antibodies.

To the Editor:— In reply to a recent query concerning the use of A and B group substances to render group O blood safe for use for recipients of all blood groups, the statement was made that there is still no direct evidence establishing the efficacy of the group substances in vivo, when used in this way (The Journal, Nov. 17, 1945, p. 840). Our purpose in this communication is to describe briefly some recent experiments which supply this direct evidence. Group B plasma having a very high titer of anti-A agglutinins was obtained from a blood donor who had been immunized by an injection of soluble group substances. The titer of the anti-A agglutinin for A 2 cells by the well slide method reached as high as 2,500 units (equivalent to 10,000 units by the tube technic or about two hundred times the average normal titer) but had fallen

SEVERAL observers1 2 3 4 have reported cases of acute anemia occurring in patients with primary atypical pneumonia who have developed high titers of cold agglutinins (autohemagglutinins). Since all these patients, with 2 possible exceptions, had been given one of the sulfonamide drugs, an additional etiologic factor was present that these authors thought might have been the primary or a contributory cause of the hemolytic crisis.† In the case reported below, no sulfonamide drug or any other medicinal agent was used that could have been an etiologic factor responsible for the causation of the acute hemolytic anemia. Case Report The patient, a 34-year-old . . .

There are two critical stages in the retroviral reprogramming of somatic cells to produce human induced pluripotent stem cell (hiPSC) lines. One is the production of high titer virus required to reprogram somatic cells; the other is identification of true hiPSC colonies from heterogeneous cell populations, and their isolation and expansion to generate a sustainable, pluripotent stem cell line. Here we describe simple, time-saving methods to address the current difficulties at these two critical junctures. First, we have developed a method to increase the number of infectious viral units 600-fold. Second, we have developed a TRA-1-81-based positive selection column method for isolating “true” hiPSCs from the heterogeneous cell populations, which overcomes the labor-intensive and highly subjective method of manual selection of hiPSC colonies. We have used these techniques to produce 8 hiPSC lines from human fibroblasts and we believe that they are of considerable utility to researchers in the hiPSC field.

SummaryHerein is described a simple, economical and quick method for preparing highly specific, high titer guinea pig immune sera for use as positive controls, and for standardizing antigens in the complement fixation test. This method has been used for producing sera for Western and Eastern equine, St. Louis, Japanese B, West Nile, Russian Spring-summer andthe Hammon-Reeves California virus.

There are multiple modalities by which trauma occurs to the neck. One of these includes minor suction trauma which usually results in a superficial bruising of the skin. While this usually self-resolves, patients with hemophilia are at higher risk for the development of bleeding from such trauma. Hematomas of the head and neck in patients with hemophilia have seldom been reported. We report a unique case of expanding bilateral neck hematomas secondary to suction trauma in a patient with Hemophilia A with high-titer inhibitor and highlight the importance of a multidisciplinary approach in the management of this complex patient.

The recent emergence of the pandemic H1N1 viruses and their being labeled as ‘swine flu’ has had several detrimental effects on the pork industry. Novel H1N1 strains have been recently detected in swine populations in the United States and in other parts of the world. While the public health significance of these findings is unknown, it is important to determine whether existing vaccines or exposure to previously circulating strains of swine influenza will protect pigs against the novel H1N1. Using a partial two-way cross-hemagglutination inhibition (HI) assay, we have determined a swine H1N1 strain present in a commercial vaccine cross-reacted with antiserum specific to the novel H1N1. We have also shown that a field serum sample with a HI high titer to a ’99 H1N1 strain crossreacted strongly with the novel H1N1. Anti-sera which were specific to the non-pandemic H1N1 strains also crossreacted with the novel H1N1. Therefore, we conclude that current vaccines and circulating non-pandemic H1N1 field strains will provide at least partial protection against the novel H1N1 virus in pigs.

This article evaluates a new and rapid group B streptococcus antigen detection method called ICON Strep B (Hybritech, Inc, San Diego), an enzyme immunoassay that uses immunoconcentration techniques. Results were compared with those of a latex particle agglutination (LPA) test and culture. ICON sensitivity for cervical and gastric aspirate samples was 92.3% and 100%, respectively, compared with culture. Specificity was 99.2% and 98.9%. Sensitivity and specificity for urine were 100% and 99.3%, respectively, compared with LPA. Accuracy of the ICON test, based on 88J specimens, was 99.2%. There was excellent agreement with both LPA and culture.

Hemagglutinating and acetylesterase functions as well as the 124 kDa glycoprotein were present in the highly cell-culture adapted, avirulent bovine coronavirus strain BCV-L9, in the Norden vaccine strain derived from it, and in 5 wild-type, virulent strains that multiplied in HRT-18 cells but were restricted in several types of cultured bovine cells. The BCV-L9 and the wild-type strain BCV-LY-138 agglutinated chicken and mouse erythrocytes. The acetylesterase facilitated break-down of the BCV-erythrocyte complex with chicken but only to a minimal extent with mouse erythrocytes in the receptor-destroying enzyme test. Purified preparations of the vaccine and the wild-type strains agglutinated chicken erythrocytes at low titers and mouse erythrocytes at 128 to 256 times higher titers whereas receptor destroying enzyme activity was detectable only with chicken erythrocytes. When wild-type strains were propagated in HRT cells at low passage levels, they produced 5 x 10(5) to 4.5 x 10(6) plaque forming units per 50 microliters which agglutinated erythrocytes from mice but not from chickens. Diisopropylfluoro-phosphate moderately increased the hemagglutination titers, but completely inhibited the receptor destroying enzyme of purified virus of all strains. It had virtually no influence on the plaque-forming infectivity of the different BCV strains. The acetylesterase of strain BCV-L9 reacting in the receptor-destroying enzyme test was stable for 3 h at 37 and 42 degrees C. It was inactivated within 30 min at 56 degrees C while the hemagglutinin function of this strain was stable for 3 h at 37, 42, and 56 degrees C, but it was inactivated at 65 degrees C within 1 h.

Two rapid group B streptococcal antigen tests were compared with nonselective blood agar culture in 1062 unselected patients admitted to labor and delivery. Vaginal specimens taken from each patient on admission were used to perform each of two rapid tests and corresponding cultures. The rapid tests were the Streptex latex agglutination assay and the Equate Strep B test, which uses a solid-phase immunoassay. Overall, 105 patients (9.9%) had at least one positive culture. The sensitivities for the rapid tests were 15.1% for Streptex and 21.5% for Equate. Specificities were 99.3 and 98.7%, respectively. Sensitivity was minimally increased in the setting of ruptured membranes for both tests. Likewise, use of separate swabs for streaking the culture plate and performing the rapid test increased the sensitivity, but this was not significant for either test. In control experiments, the limit of sensitivity of both rapid tests was 5 x 10(6) colony-forming units. We conclude that at present, these tests are not sensitive enough for routine use in this type of clinical setting.

This study is a cross sectional and descriptive study. Sample population was the women referred for marriage consultation to Gorgan Marriage Consultation Center in 2004. The prevalence of toxoplasma antibodies (IgG, IgM) was determined by ELISA method. Blood samples were collected randomly from 300 women referred to the center for consultation. They were transferred to the laboratory for antibodies determination. Gorgan located in north of Iran and south east of Caspian sea. The results showed that the general prevalence to the positive cases based on high titer of IgG was 48.3% and of IgM was 11.7%. There was not any relation between positive cases and age, education, place of residence, job, keeping domestic animals (except cat) vegetable consumption and wash-up, but there was a relation between the positive cases of IgM and keeping cat at home (PV

Thrombotic thrombocytopenic purpura (TTP) is rarely associated with undifferentiated connective tissue disorder. We present a patient with TTP, undifferentiated connective tissue disorder, and very high titer of anti-ribonucleoprotein antibodies. Her TTP did not respond to intravenous methylprednisolone and cyclophosphamide requiring her to remain dependent on plasmapheresis. Her disease remitted successfully after 4 doses of rituximab given at weekly intervals and she remained in continuous remission 6 months after therapy. We propose early and aggressive use of B-cell depletion therapy in TTP associated with autoimmune disorders.

Paraneoplastic optic neuropathy (PON) is a rare syndrome usually associated with small cell lung carcinoma. In the 27 rigorously reported cases, neurologic manifestations other than visual loss have been present in all but 2. In the single case in which vision improved in response to treatment of the cancer, the collapsin response-mediating protein (CRMP)-5 titer did not change, and the ophthalmic examination was not detailed. We describe a patient with optic neuropathy and vitritis as the only clinical manifestations of PON marked by an extremely high titer of CRMP-5 antibody. Treatment of the underlying small cell lung cancer coincided with resolution of the visual abnormalities and a dramatic decrease in the CRMP-5 titer.

Idiopathic inflammatory myopathies such as polymyositis (PM) and dermatomyositis (DM) are a group of rare autoimmune diseases, characterized by an inflammatory infiltrate within the skeletal muscle and high titer of circulating autoantibodies in the patient's serum. The etiopathogenesis of these diseases is not known and the relationship between the specific muscle involvement and the ubiquitary presence of the targeted antigens is still unclear. The enhanced expression of myositis specific autoantigens in regenerating muscle fibers from biopsies of PM and DM patients compared to normal muscle has been recently demonstrated. In order to understand whether candidate autoantigens in myositis are expressed during post-natal myogenesis, we performed immunolocalization studies of myositis specific autoantigens in skeletal muscle from newborn and adult rats. Our observations indicate the presence of myositis specific autoantigens during post-natal myogenesis, with possible implications for the induction and/or amplification of the immune-inflammatory response, in patients affected with autoimmune myositis.

Prophylactic therapy in patients with hemophilia A and B is well known and established treatment today. Typically, prophylactic therapy in hemophilia A patients is given in a dose of 25–40 IU/kg three times per week. This dosing targets a pre infusion level of > 1% in order to mimic the clinical phenotype of moderate hemophilia. Follow up studies of patients with prophylactic therapy since early childhood demonstrated a significantly reduced prevalence of joint abnormalities and disableness compared with patients treated with FVIII only on-demand. Patients with hemophilia A and inhibitors usually have more pronounced joint disease in compare with patients without inhibitor and the bleeding episodes are harder to treat. It has been already published the possibility to introduce prophylactic therapy in patients with hemophilia and inhibitors with bypassing agent.

Acquired hemophilia is a rare severe bleeding disorder caused by antibodies against coagulation factors. Factor VIII is the commonest factor affected.

During a 5-month period, 248 general practitioners from 164 general practice offices obtained duplicate throat swabs from 2469 patients with acute pharyngotonsillitis. At the general practice office, one of the swabs was assayed for the presence of group A streptococcal antigen, using one of five commercially available diagnostic kits. (The participants did not receive instruction in the use of the kits.) The other swab was cultured in our laboratory for the presence of group A streptococci. The diagnostic parameters for the five kits were as follows: Abbott TestPack Strep A—sensitivity, 79% ( 110 139 ) and specificity, 98% ( 490 501 ); Direct Strep A EIA (Roche)—sensitivity, 79% ( 104 132 ) and specificity, 63% ( 178 281 ); Respirastick—sensitivity, 55% ( 46 84 ) and specificity, 96% ( 298 309 ); Reveal Colour Strep A—sensitivity, 82% ( 114 139 ) and specificity, 83% ( 223 268 ); Tandem ICON Strep A—sensitivity, 78% ( 111 143 ) and specificity, 98% ( 362 368 ). The antigen detection test results were heavily influenced by the clinical prediction of a streptococcal or viral etiology. It is concluded that in the absence of a standard throat culture, the Abbott TestPack Strep A kit and the Tandem ICON Strep A kit both offer a more safe diagnostic alternative than clinical assessment. Depending on the current prevalence of group A streptococci, the additional use of a throat swab culture should be considered in case of negative antigen detection test results.

Summary Following oral inoculation, the excretion of a virulent or an attenuated strain of bovine coronavirus in calf feces and their persistence in spiral colon, ileum, and jejunum were studied using hemagglutination and direct fluorescent antibody tests, respectively. The virus was excreted in feces for only 3 days at high titers and then intermittently at low titers. However, viral antigen was detected in crypt or Peyer's patches for at least 3 weeks after infection in 1 of 3 calves given virulent virus and 1 of 2 calves given attenuated virus.

Previously, we reported that the sensitivity of plasma DNA for patients with pulmonary emboli was 83 to 88 percent. To confirm these findings in a more comprehensive study, we collected plasma samples from 137 consecutive patients undergoing 148 ventilation-perfusion lung scans for pulmonary embolism. DNA was measured using a counter-immunoelectrophoresis technique that used high titer precipitating double-stranded DNA antibody from a patient with systemic lupus erythematosus. In addition to 17 patients (17 lung scans) excluded for not having plasma collected, 32 patients (37 lung scans) were excluded for having either a condition other than pulmonary embolism that could be associated with plasma DNA or for having nonacute symptoms. Eighteen of 22 patients with a diagnosis of pulmonary embolism (defined by either a high probability lung scan or abnormal pulmonary angiogram) had detectable plasma DNA. Only four of 27 patients without pulmonary embolism (defined by either a normal lung scan or normal pulmonary angiogram) had plasma DNA detected. Based on these results, plasma DNA had a sensitivity of 82 percent and a specificity of 85 percent for this condition. Plasma DNA is a promising test for pulmonary embolism and could help physicians interpret equivocal lung scan findings and thereby clarify difficult decisions such as the need for pulmonary angiography. (Chest 1990; 97:63-68)

The purpose of this study was to evaluate the overall performance of rapid antigen detection (RAD) in group A streptococcus (GAS) in Finland by using the results of external quality assurance (EQA) samples. We also compared the performance of laboratory professionals to that of nursing professionals. Around 22,800 EQA results among a total of 383 laboratories and physician’s offices were analysed. Vocational data on the personnel who carried out the tests were available for 10,088 EQA samples, 7,428 of which were tested by laboratory technicians and 2,531 by nursing staff. The best overall performance was found with GAS-negative samples: 99% of the reports were correct. In contrast, the overall performance was only 76% when the samples were weakly positive for GAS antigen. The laboratory technicians performed statistically significantly better than the nursing staff, with both strongly positive (correct results 98.9% vs. 95.1%, respectively; p

Lentiviral vectors offer unique versatility and robustness as vehicles for gene delivery. They can transduce a wide range of cell types and integrate into the host genome in both dividing and post-mitotic cells, resulting in long-term expression of the transgene both in vitro and in vivo. This protocol describes how lentiviral vectors can be produced, purified and titrated. High titer suspensions can be routinely prepared with relative ease: a low-titer (10(6) viral particles/ml) unpurified preparation can be obtained 3 d after transfecting cells with lentiviral vector and packaging plasmids; a high-titer (10(9) viral particles/ml) purified preparation requires 2 more days.