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Descreve-se um caso de amebíase hepática num homem de 41 anos de idade e discutem-se as dificuldades diagnósticas, nomeadamente o significado da positividade da reacção de Widal para S. paratyphi B. Chama-se a atenção para o quadro clínico dominado por síndrome febril e hepatomegalia dolorosa, espontaneamente e a percussão, e revem-se as características clinicas, laboratoriais e radiológicas do abcesso hepático amebiano. Discute-se a terapêutica instituída com Metronidazol, notável pela sua elevada eficácia e ausência de toxicidade. Realça-se a necessidade do diagnóstico precoce desta afecção e a sua importância crescente entre nos, dado o elevado número de indivíduos residentes em Portugal, provenientes de regiões onde a amebíase é endémica.

Melioidosis is an infection caused by the bacterium Burkholderia pseudomallei. The most common presentation is bacteremia occurring in 38–73% of all patients, and the mortality rate ranges from 9% to 42%. Although there is abundant data representing risk factors for infection and patient outcomes, there is limited information regarding laboratory investigations associated with bacteremia and mortality. We assessed a range of baseline and diagnostic investigations and their association with patient outcomes in a retrospective cohort study in Townsville, Australia. 124 patients’ medical and laboratory records were reviewed between January 1, 1997 and December 31, 2020. Twenty-seven patients died and 87 patients were bacteremic. The presence of lymphopenia (< 1.5 × 109 cells/L) was the highest risk for bacteremia (relative risk [RR] 2.2; 95% CI: 1.3–3.7, P < 0.001). Factors associated with mortality included lymphopenia, (RR: 1.4; 95% CI: 1.2–1.6, P = 0.004); uremia (RR: 1.7; 95% CI: 1.1–2.5, P = 0.03); and an elevated international normalized ratio (RR: 1.5; 95% CI: 1.2–2.0, P = 0.006). Median incubation to positive blood culture result was 28 hours with 15/82 (18%) positive in ≤ 24 hours. For serological testing during admission only 53/121 (44%) were indirect hemagglutination assay positive, 67/120 (56%) enzyme immunoassay IgG positive, and 23/89 (26%) IgM positive. Simple baseline investigations at time of presentation may be used to stratify patients at high risk for both bacteremia and mortality. This information can be used as a decision aid for early intensive management.

For many years natural remedies have played a significant part in pharmaceutical biology. Even today, many health organizations depend on the natural products, WHO estimated that more than 80% population depends on the natural remedies. Chia and basil seeds are herbs that contains several beneficial minerals, proteins, vitamins and dietary fibers. They also contain omega 3 and omega 6 fatty acids (healthy) as well as the sufficient amount of polyphenols and antioxidants, like caffeic acids, myricetin and others. Current study consists of comparing the Total Antioxidant Capacity (TAC) of Chai and basil seeds with the help of various assays. Confirmatory tests to verify the presence of phytolectins in both chia and basil seeds have been done by performing hemagglutination tests. These phytolectins are the proteins that binds to the oilgosaccharides and monosaccharides in the reversible manner. Although further studies are required for isolating and characterizing the phytolectins from the seeds.

Introduction: Cognitive impairment has been reported in people living with HIV-1 (PLWH) with prior syphilis, while PLWH who present with incident syphilis have reduced blood CD4 T-lymphocyte and elevated HIV-1 RNA levels. However, the clinical, virological and neurocognitive effects of syphilis during acute HIV-1 (AHI) remain unknown. Methods: Pre-antiretroviral therapy laboratory outcomes and neurocognitive performance in a four-test battery in the SEARCH10/RV254 AHI cohort were compared according to syphilis status, determined by serum Treponema pallidum haemagglutination (TPHA), Venereal Disease Research Laboratory (VDRL) and syphilis treatment history. Impaired cognitive performance was defined as having z-scores ≤ −1 in at least two tests or ≤ −2 in at least one test. Results: Out of 595 AHI participants (97% male, median age of 26 years and estimated duration of HIV-1 infection of 19 days), 119 (20%) had history of syphilis (TPHA-positive), of whom 51 (9%) had untreated syphilis (TPHA-positive/VDRL-positive/without prior treatment). Compared with those without syphilis (TPHA-negative), individuals with untreated syphilis had higher CD8 T-lymphocyte levels but not higher plasma HIV-1 RNA or lower CD4 T-lymphocyte levels. Taking into account estimated duration of HIV-1 infection (P

Cystic echinococcosis (CE) is a common public health concern in Turkey. In this study, we investigated the agreement between the results of radiological imaging methods and serological tests through a 5-year retrospective evaluation in patients admitted to a university hospital with a suspicion of CE so as to determine the frequency of CE in the study region.The indirect hemagglutination test (IHA) results of 1.046 patients obtained from various clinics with the suspicion of CE between January 2014 and December 2018 were retrospectively analysed. Of these, patients with at least one radiological imaging report in the system (938 patients) were included in the study. Radiological imaging findings and IHA test results were compared and examined.Seropositivity was detected by IHA test in 143 (15.2%) of 938 patients included in the study. The CE findings were recorded in at least one radiological imaging report in 130 (90.9%) of 143 patients with positive IHA test. At least one of the radiological imaging reports suggested presence of CE in 362 (38.5%) of all the patients.Thus, serological test and radiological imaging methods should be used in combination for the diagnosis of CE.Kistik ekinokokkoz (KE) ülkemizde yaygın görülen bir halk sağlığı problemidir. Bu çalışmada bir üniversite hastanesinde geriye dönük beş yıllık dönemde, KE şüphesiyle istenilen radyolojik görüntüleme yöntemleri ile serolojik test sonuçları arasındaki uyumun araştırılması ve yaşadığımız bölgedeki KE sıklığının değerlendirilmesi amaçlanmıştır.Çalışmada Ocak 2014 ile Aralık 2018 tarihleri arasında KE şüphesi ile çeşitli kliniklerden gönderilen 1,046 hastanın indirekt hemaglütinasyon test (IHA) sonuçları retrospektif olarak incelenmiştir. İncelenen 1,046 hastadan sistemde kayıtlı en az bir radyolojik görüntüleme raporu olduğu tespit edilen 938’i bu çalışmaya dahil edilmiştir. Radyolojik görüntüleme bulguları ve IHA test sonuçları irdelenmiştir.Çalışmaya alınan 938 hastanın 143’ünde (%15,2) IHA testi ile seropozitiflik saptanmıştır. IHA testi pozitif bulunan 143 hastanın 130’unda (%90,9) en az bir radyolojik görüntüleme raporunda KE bulgusu olduğu kaydedilmiştir. Tüm hastaların ise 362’sinde (%38,5) radyolojik görüntüleme raporlarından en az birinde KE bulgusu görülmüştür.Çalışmada KE tanısında serolojik test ve radyolojik görüntüleme yöntemlerinin birlikte kullanılması gerektiği sonucuna varılmıştır.

Prediction models indicate that melioidosis may be common in parts of East Africa, but there are few empiric data. We evaluated the prevalence of melioidosis among patients presenting with fever to hospitals in Tanzania. Patients with fever were enrolled at two referral hospitals in Moshi, Tanzania, during 2007–2008, 2012–2014, and 2016–2019. Blood was collected from participants for aerobic culture. Bloodstream isolates were identified by conventional biochemical methods. Non–glucose-fermenting Gram-negative bacilli were further tested using a Burkholderia pseudomallei latex agglutination assay. Also, we performed B. pseudomallei indirect hemagglutination assay (IHA) serology on serum samples from participants enrolled from 2012 to 2014 and considered at high epidemiologic risk of melioidosis on the basis of admission within 30 days of rainfall. We defined confirmed melioidosis as isolation of B. pseudomallei from blood culture, probable melioidosis as a ≥ 4-fold rise in antibody titers between acute and convalescent sera, and seropositivity as a single antibody titer ≥ 40. We enrolled 3,716 participants and isolated non-enteric Gram-negative bacilli in five (2.5%) of 200 with bacteremia. As none of these five isolates was B. pseudomallei, there were no confirmed melioidosis cases. Of 323 participants tested by IHA, 142 (44.0%) were male, and the median (range) age was 27 (0–70) years. We identified two (0.6%) cases of probable melioidosis, and 57 (17.7%) were seropositive. The absence of confirmed melioidosis from 9 years of fever surveillance indicates melioidosis was not a major cause of illness.

Innate immunity is vital for the early control of influenza A virus (IAV) infection. Serum amyloid A (SAA1) is an acute phase reactant produced in the liver and lung that rises dramatically during IAV infection. The potential role of SAA1 in host defense against IAV is unknown. SAA1 has been reported to directly activate neutrophils and to recruit them to the lung during infectious and inflammatory processes. Neutrophils are the most abundant cell recruited to the lung in the early phase of IAV infection. There are different forms and preparations of SAA1 that have found to have different effects on phagocyte responses, through various receptors. In this paper, we test the direct effects of various preparations of serum derived or recombinant SAA on IAV and how it modulates the interactions of IAV with neutrophils. All SAA preparations bound to IAV in vitro but caused minimal hemagglutination inhibition or viral aggregation. The human serum‐derived SAA1 or the complex of SAA1 with HDL did have IAV neutralizing activity in vitro, whereas the recombinant SAA1 preparations did not. We found that different SAA preparations also had markedly different effects on neutrophil functions, with E. coli‐derived SAA1 triggering some responses in neutrophils on its own or in presence of IAV whereas mammalian cell‐derived SAA1 did not. This discrepancy could be explained by the reported contamination of the former preparation with bacterial components. Of interest, however, serum SAA alone, serum SAA complexed with HDL, or HDL alone potentiated some neutrophil responses to IAV. Our results suggest that SAA may play some role in host response to IAV, but further work needs to be done to clarify the role of different variants of SAA alone or complexed with HDL., Graphical Abstract Serum amyloid a protein (SAA) bound to and inhibited influenza virus infectivity, but SAA free of bacterial products did not alter neutrophil responses.

Urinary bladder cancer is a common malignancy in Egypt, thus reliable methodologies are required for screening and early detection. In this study, we analyzed the gene expression of a Schistosoma hematobium specific microRNA “Sha-miR-71a” and mitogen-associated protein kinase-3 (MAPK-3) in the urine samples of 50 bladder cancer patients and 50 patients with benign bilharzial cystitis. Fifty control subjects were also tested. Indirect hemagglutination test (IHA) diagnosed 70% of studied cancer cases as bilharzial associated bladder cancer (BBC), while histopathological examination detected only 18%. Urinary Sha-miR-71a & MAPK-3 revealed enhanced expression in BBC (p-value = 0.001) compared to non-bilharzial bladder cancer (NBBC) cases. Patients with chronic bilharzial cystitis exhibited a significant increase in gene expression compared to those with acute infection (p-value = 0.001). Sha-miR-71a and MAPK-3 showed good sensitivity and specificity in the diagnosis of BBC when analyzed by the receiver operating characteristic (ROC) curve. They were also prognostic regarding malignancy grade. Both biomarkers showed a positive correlation. Our results revealed that IHA is a reliable test in the diagnosis of bilharziasis associated with bladder cancer, and that Sha-miR-71a and MAPK-3 provide non-invasive specific biomarkers to diagnose BBC, as well as a potential role in testing bilharzial patients for risk to develop cancer.

Introduction Human cystic echinococcosis (CE) diagnosis is based on imaging findings and serology. Serology is the first-line test followed by imaging tests. Materials and methods A total of 268 serum samples from CE patients were included in this study. The serodiagnosis was made simultaneously by indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), immunoblotting test (IB), and immunoelectrophoresis (IEP). Results In serum samples belonging to surgically confirmed CE patients, we observed a percentage of positivity of 83.7% [77.2%–89.0%] and 80.7% [73.9%–86.4%] for IHA and IgG-ELISA, respectively. IgG-IB was associated with a higher positivity rate than IEP with 81.3% [74.6%–86.9%] and 62.0% [54.2%–69.5%], respectively. IHA and ELISA results were analyzed using cut-off generated by receiver operating curves. The best diagnostic performances were achieved by IHA (cut-off ≥ 1/128) and ELISA (> 1.16 index). Sensitivities reported in patients with suggestive imaging findings and positive Echinococcus IB were 86.2% [78.0%–92.2%], 72.5% [62.8%–80.9%], 49.0% [39.0%–59.1%] for IHA, IgG-ELISA, and IEP, respectively. All tests gave false negative results in the confirmed CE group. Overall, 18.6% of negative results were obtained by IgG-IB test. Cross-reactivities with non-hydatid serum samples were observed in all tests. Only one patient carrying Taenia saginata serum cross-reacted with 8/12 kDa band by IB. We observed specificity at 73% [63.2%–81.4%], 87% [78.8%–92.9%], 99% [94.6%–100.0%], and 99% [94.6%–100.0%] with IHA, IgG-ELISA, IEP, and IgG-IB, respectively. Serology was less sensitive (74%) in lung cysts. Sensitivity was better in liver cysts, especially by IgG-IB (96%).

The H7N9 viruses have been circulating for six years. The insertion of a polybasic cleavage site in the haemagglutinin (HA) protein of H7N9 has resulted in the emergence of a highly pathogenic (HP) avian influenza virus. Currently, there are limited studies on neutralizing monoclonal antibodies(mAbs) against HP H7N9 AIVs. In this study, mice were immunized with inactivated H7N9 vaccine of A/ZJU01/PR8/2013 to produce murine mAbs. Finally, two murine mAbs against the HA of low pathogenic (LP) virus were produced and characterized. Characterization included determining mAbs binding breadth and affinity, in vitro neutralization capacity, and potential in vivo protection. Two of these mAbs, 1H10 and 2D1, have been identified to have therapeutic and prophylactic efficacy against the HP strain in mouse passive transfer-viral challenge experiments. The mAb 1H10 was most efficacious, even if the treatment-time was as late as 72 h post-infection, or the therapeutic dose was as low as 1 mg/kg; and it was confirmed to have haemagglutination inhibition and neutralizing activity on both LP-and HP-H7N9 strains. Further study indicated that the protection provided by 2D1 was mediated by antibody-dependent cellular cytotoxicity. The mAbs described here provide promising results and merit further development into potential antiviral therapeutics for H7N9 infection.

Aim: Diagnosis of echinococcosis is based on clinical symptoms, imaging techniques and particularly on the detection of specific antibodies in serum specimens of patients. The aim of this study is retrospective evaluation of seropositivity from patients who is diagnosed as Cystic echinococcosis. Materials and Methods: In this study, the blood serum samples of patients, which were sent to the Medical Microbiology Laboratory from several clinics of the Selcuk University Medical Faculty Hospital between 01 January 2015- 01 July 2020, were evaluated in terms of Cystic echinococcosis by using indirect hemaglutination test. The results were evaluated with a cut off titer ≥160 of as recommended by the manufacturer. Results were evaluated by gender and ages. Results: Cystic echinococcosis (CE) was determined to be seropositive 332 (21.6%) of 1543 patient samples. Seropositivity was found in 226 (14,7%) of 933 female patients and in 106 (6,9%) of 610 male patients. Seropositivity was mostly seen in 21-40 age group. Conclusion: While there have been a number of animal studies, little information is available about the prevalence of Cystic echinococcosis in humans in Turkey. Cystic echinococcosis continues to be a majör public health problem in the Konya province. Large scale prevention and control programs should be implemented against this disease.

This study aimed to retrospectively examine the indirect haemagglutination (IHA) test results of patients suspected of cystic echinococcosis (CE) and admitted to Bursa Uludağ University Health Practice and Research Center Hospital.Serum samples in the serology laboratory of our hospital were evaluated using the commercial CellognostSeropositivity was determined in 213 (19.9%) of 1.072 patients suspected of having CE by the IHA method. Of the patients with seropositivity, 120 (56.3%) were female and 93 (43.7%) were male. The highest positivity rate in both sexes was found in patients aged 20-29 years (22.5% in women; 14.1% in men).The results indicate that CE maintains its importance as a public health problem in Bursa as in Turkey.Bursa Uludağ Üniversitesi Sağlık Uygulama ve Araştırma Merkez Hastanesi’ne kistik ekinokokkoz (KE) şüphesiyle başvuran hastalarda indirekt hemaglütinasyon (IHA) test sonuçlarının geriye yönelik olarak incelenmesi amaçlanmıştır.Merkezimizin seroloji laboratuvarında serum örnekleri ticari CellognostÇalışmada KE şüpheli 1,072 hastanın 213’ünde (%19,9) IHA yöntemi ile seropozitiflik saptanmıştır. Seropozitiflik saptanan hastaların 120’si (%56,3) kadın, 93’ü (%43,7) erkek olarak belirlenmiştir. Her iki cinstede en fazla pozitiflik oranı 20-29 yaş grubunda (kadınlarda %22,5, erkeklerde %14,1) grubunda elde edilmiştir.KE’nin ülkemizde olduğu gibi ilimizde de halk sağlığı sorunu olarak önemini koruduğu sonucuna varılmıştır.

Background and objectives To detect HPA-15 alloantibodies, we previously developed a human platelet antigen 15 (HPA-15)-expressing cell line-based modified rapid monoclonal antibody immobilization of platelet antigen (CL-MR-MAIPA) assay. In this study, the protocol was modified for easier performance by introducing the mixed-passive haemagglutination (MPHA) principle. Material and methods In total, 20 samples that tested negative for HPA alloantibodies and eight that tested positive for HPA-15 alloantibodies (two and six positive for HPA-15a and HPA-15b antibodies, respectively) by CL-MR-MAIPA assay were used in this study. HPA-15 cell lines were incubated with serum/plasma and then solubilized. The lysate was transferred to a round-bottom well, which was coated with anti-human CD109 monoclonal antibodies. After incubation and repeated washings, sheep red blood cells, coated with anti-human IgG, were added to the wells. Haemagglutination was assessed the next day. Results The proposed cell line-based immune complex capture-dependent mixed-passive haemagglutination (CL-IC-MPHA) assay consisted of four steps, but required only 2 h to perform, except for overnight incubation for haemagglutination. Two HPA-15a alloantibody samples were reactive only for HPA-15a cells, and six HPA-15b alloantibody samples were reactive only for HPA-15b cells with the CL-IC-MPHA assay. The 20 samples that tested negative for HPA alloantibodies did not react with HPA-15a or HPA-15b cells. These data indicated that the CL-IC-MPHA assay was highly specific and sensitive. Unfortunately, the CL-IC-MPHA assay's analytic sensitivity was twofold to eightfold lower than that of the CL-MR-MAIPA assay. Conclusion A novel, easy-to-perform protocol was successfully developed to detect HPA-15 alloantibodies with high specificity and sensitivity.

Agaricus bisporus mannose binding protein (Abmb) demonstrates permeability to epithelial monolayer barrier of the intestine, resistance to gastrointestinal tract conditions and to proteolysis therefore it holds potential as a drug carrier for oral route administration. Abmb also display antiproliferative activity to breast cancer cells and stimulation of immune system thus could potentially be also developed for therapeutic purpose. It is not immunogenic or toxic thereby safe for use. In this paper we further provide evidence that Abmb also lacks of agglutinating activity despite sharing high structural homology to lectins. Abmb is thereby the only mannose specific binding protein that is not member of lectin family. This evidence provides further support on the use of Abmb as pharmaceutical or medicinal agent. Its molecular globularity that may contribute to its lack of agglutination capacity was also evaluated.

Toxoplasma gondii is the pathogen of toxoplasmosis, which affects a wide range of animal species and humans. In the present study, a serological investigation was undertaken to determine the seroprevalence of T. gondii infection in free-ranging pigs in northeastern China. A total of 806 serum samples were collected from free-ranging pigs in 6 cities in Liaoning, Jilin, and Heilongjiang Provinces. Among them, 163 serum samples (20.22%) were detected T. gondii positive by indirect hemagglutination test (IHA). In each administrative district, the prevalence ranges from 12.99 to 22.22%. It was found that sample collection year and season were associated with T. gondii infection. Samples collected in 2015 and 2016 had had higher positive rate compared to those collected in 2017. Samples collected in summer and autumn also have higher positive rate than those in winter. This is the first report on T. gondii seroprevalence in free-ranging pigs in northeastern China, which adds the epidemiological data of T. gondii infection in free-ranging pigs in China. These results indicated that free-ranging pigs in Liaoning, Jilin, and Heilongjiang Provinces are exposed to T. gondii, which is a significant threat to local pig industry as well as public health. It is necessary to take relevant comprehensive measures to prevent and control T. gondii infection in free-ranging pigs in these regions.

Serologic, point-of-care tests to detect antibodies against SARS-CoV-2 are an important tool in the COVID-19 pandemic. The majority of current point-of-care antibody tests developed for SARS-CoV-2 rely on lateral flow assays, but these do not offer quantitative information. To address this, we developed a new method of COVID-19 antibody testing employing hemagglutination tested on a dry card, similar to that which is already available for rapid typing of ABO blood groups. A fusion protein linking red blood cells (RBCs) to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein was placed on the card. 200 COVID-19 patient and 200 control plasma samples were reconstituted with O-negative RBCs to form whole blood and added to the dried protein, followed by a stirring step and a tilting step, 3-minute incubation, and a second tilting step. The sensitivity for the hemagglutination test, Euroimmun IgG ELISA test and RBD-based CoronaChek lateral flow assay was 87.0%, 86.5%, and 84.5%, respectively, using samples obtained from recovered COVID-19 individuals. Testing pre-pandemic samples, the hemagglutination test had a specificity of 95.5%, compared to 97.3% and 98.9% for the ELISA and CoronaChek, respectively. A distribution of agglutination strengths was observed in COVID-19 convalescent plasma samples, with the highest agglutination score (4) exhibiting significantly higher neutralizing antibody titers than weak positives (2) (p

The COVID-19 pandemic has caused significant morbidity and mortality. There is an urgent need for serological tests to detect antibodies against SARS-CoV-2, which could be used to assess past infection, evaluate responses to vaccines in development, and determine individuals who may be protected from future infection. Current serological tests developed for SARS-CoV-2 rely on traditional technologies such as enzyme-linked immunosorbent assays (ELISA) and lateral flow assays, which have not scaled to meet the demand of hundreds of millions of antibody tests so far. Herein, we present an alternative method of antibody testing that depends on one protein reagent being added to patient serum/plasma or whole blood with direct, visual readout. Two novel fusion proteins, RBD-2E8 and B6–CH1-RBD, were designed to bind red blood cells (RBCs) via a single-chain variable fragment (scFv), thereby displaying the receptor-binding domain (RBD) of SARS-CoV-2 spike protein on the surface of RBCs. Mixing mammalian-derived RBD-2E8 and B6–CH1-RBD with convalescent COVID-19 patient serum and RBCs led to visible hemagglutination, indicating the presence of antibodies against SARS-CoV-2 RBD. B6–CH1-RBD made in bacteria was not as effective in inducing agglutination, indicating better recognition of RBD epitopes in mammalian cells. Given that our hemagglutination test uses methods routinely used in hospital clinical labs across the world for blood typing, we anticipate the test can be rapidly deployed in low-resource settings at minimal cost. We anticipate our hemagglutination assay may find extensive use in low-resource settings for detecting SARS-CoV-2 antibodies.

Herbal medicines are becoming more popular and acceptable day by day due to their effectiveness, limited side effects, and cost-effectiveness. Cholistani plants are reported as a rich source of antibacterial, antifungal, antiprotozoal, antioxidant, and anticancer agents. The current study has evaluated antiviral potential of selected Cholistani plants. The whole plants were collected, ground and used in extract formation with n-hexane, ethyl acetate and n-butanol. All the extracts were concentrated by using a rotary evaporator and concentrate was finally dissolved in an appropriate vol of the same solvent. All of the extracts were tested for their antiviral potential by using 9-11 days old chick embryonated eggs. Each extract was tested against the Avian Influenza virus H9N2 strain (AIV), New Castle Disease virus Lasoota strain (NDV), Infectious bronchitis virus (IBV) and an Infectious bursal disease virus (IBDV). Hemagglutination test (HA) and Indirect Hemagglutination (IHA) tests were performed for different viruses. The overall order of the antiviral potential of Cholistani plants against viruses was NDV>IBV>IBDV>AIV. In terms of antiviral activity from extracts, the order of activity was n-butanol>ethyl acetate>n-hexane. The medicinal plants Achyranthes aspera, Neuroda procumbens, Panicum antidotale, Ochthochloa compressa and Suaeda fruticose were very effective against all four poultry viruses through their extracts. The low IC50 values of these extracts confirm the high antiviral potential against these viruses. It is worth to mention that Achyranthes aspera was found positive against IBDV through all its extracts which overcome the problem of unavailability of any known drug against IBDV. In short, the study proved that Cholistani plants are rich source of antiviral agent and their extracts can be used as good source of antiviral drugs both in crude and in purified form.

Since 2014, clade 2.3.4.4 has become the dominant epidemic branch of the Asian lineage H5 subtype highly pathogenic avian influenza virus (HPAIV) in southern and eastern China, while the H5N6 subtype is the most prevalent. We have shown earlier that lack of glycosylation at position 158 of the hemagglutinin (HA) glycoprotein due to the T160A mutation is a key determinant of the dual receptor binding property of clade 2.3.4.4 H5NX subtypes. Our present study aims to explore other effects of this site among H5N6 viruses. Here we report that N-linked glycosylation at site 158 facilitated the assembly of virus-like particles and enhanced virus replication in A549, MDCK, and chicken embryonic fibroblast (CEF) cells. Consistently, the HA-glycosylated H5N6 virus induced higher levels of inflammatory factors and resulted in stronger pathogenicity in mice than the virus without glycosylation at site 158. However, H5N6 viruses without glycosylation at site 158 were more resistant to heat and bound host cells better than the HA-glycosylated viruses. H5N6 virus without glycosylation at this site triggered the host immune response mechanism to antagonize the viral infection, making viral pathogenicity milder and favoring virus spread. These findings highlight the importance of glycosylation at site 158 of HA for the pathogenicity of the H5N6 viruses.

Serological detection of antibodies to SARS-CoV-2 is essential for establishing rates of seroconversion in populations, and for seeking evidence for a level of antibody that may be protective against COVID-19 disease. Several high-performance commercial tests have been described, but these require centralised laboratory facilities that are comparatively expensive, and therefore not available universally. Red cell agglutination tests do not require special equipment, are read by eye, have short development times, low cost and can be applied at the Point of Care. Here we describe a quantitative Haemagglutination test (HAT) for the detection of antibodies to the receptor binding domain of the SARS-CoV-2 spike protein. The HAT has a sensitivity of 90% and specificity of 99% for detection of antibodies after a PCR diagnosed infection. We will supply aliquots of the test reagent sufficient for ten thousand test wells free of charge to qualified research groups anywhere in the world., Serological detection of antibodies against SARS-CoV-2 can help establish rates of seroconversion. Here the authors develop a red cell agglutination test to detect antibodies against the receptor binding domain for distribution free of charge to qualified research groups.

Background While congenital syphilis is a significant public health problem that can cause severe disabilities, little is known about the situation in Uganda. We describe prevalence, associated factors and clinical presentation of congenital syphilis in Mbarara, Uganda. Methods A cross sectional study was carried out among mother- newborn dyads from the postnatal ward of Mbarara Regional Referral Hospital (MRRH). After obtaining informed consent, a structured questionnaire was used to capture data on risk factors for congenital syphilis. A finger prick was performed on the mothers for Treponema Pallidum Haemagglutination Assay (TPHA). If TPHA was positive, a venous blood sample was collected from the mother to confirm active infection using Rapid Plasma Reagin (RPR). Venous blood was drawn from a newborn if the mother tested positive by TPHA and RPR. A newborn with RPR titres 4 times higher than the mother was considered to have congenital syphilis. We fit logistic regression models to determine factors associated with congenital syphilis. Results Between June and September 2015, we enrolled 2500 mothers and 2502 newborns. Prevalence of syphilis was 3.8% (95% CI 3.1–4.6) among newborn infants and 4.1% (95% CI 3.4–5.0) among their mothers. Maternal age

Novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus (IAV), and norovirus (NV) are highly contagious pathogens that threaten human health. Here we focused on the antiviral potential of the medicinal herb, Saxifraga spinulosa (SS). Water-soluble extracts of SS were prepared, and their virus-inactivating activity was evaluated against the human virus pathogens SARS-CoV-2 and IAV, we also examined virucidal activity against feline calicivirus and murine norovirus, which are surrogates for human NV. Among our findings, we found that SS-derived gallocatechin gallate compounds were capable of inactivating all viruses tested. Interestingly, a pyrogallol-enriched fraction (Fr 1C) inactivated all viruses more rapidly and effectively than did any of the component compounds used alone. We found that 25 &micro, g/mL of Fr 1C inactivated &gt, 99.6% of SARS-CoV-2 within 10 s (reduction of &ge, 2.33 log10 TCID50/mL). Fr 1C resulted in the disruption of viral genomes and proteins as determined by gel electrophoresis, electron microscopy, and reverse transcription&ndash, PCR. Taken together, our results reveal the potential of Fr 1C for development as a novel antiviral disinfectant.

Longevity patterns in most vertebrates suggest that females benefit most from maintenance investment. A reversed longevity pattern in loggerhead musk turtles (Sternotherus minor) allowed us to test trade-offs between maintenance and survivorship. We tested the hypothesis that the sex with greater longevity has greater maintenance than the sex with shorter longevity. We also compared the following parameters between sexes: Bactericidal ability (BA) and heterophil:lymphocyte ratios (HLR). Baseline blood samples were collected from turtles in the field; a subset of turtles was returned to a laboratory for experiments of acquired immune responses to sheep red blood cells (SRBC). We found no support for the original hypothesis of reversal in sex-dependent immune trade-offs (difference between sex SRBC titers: p = .102; interaction between treatment and sex: p = .177; difference between treatments: p

Objectives Testing for drug-dependent antibodies is traditionally performed with the tube method either with drug-treated red blood cells or with untreated red blood cells in the presence of soluble drug. Gel microcolumn agglutination method was compared to tube testing for the demonstration of drug-dependent antibodies in the presence of soluble drug. Materials and methods Patient's samples were tested in parallel by tube and gel microcolumn agglutination method with untreated and/or enzyme-treated red blood cells in the presence of soluble drug. Results Twenty six different patient's samples were studied and thirty nine tests performed to investigate antibodies directed against fifteen different drugs. There was a good correlation between the results obtained by tube and gel method in terms of analytical sensitivity and specificity. Reactions appeared to be stronger with the gel test than seen with the conventional tube method for most of the drug antibodies investigated. Enzyme-treated cells should be used in addition to untreated cells to improve the sensitivity of the method for detecting drug-dependent antibodies especially those directed against drugs that do not bind firmly to red blood cells. Conclusions Gel method appeared to be sensitive, reliable, reproducible, and comparable to the conventional tube method for the detection of all the drug-dependent antibodies investigated in this study. Further studies need to be performed to evaluate gel testing for the detection of drug-dependent antibodies that only react with drug-treated red blood cells.

Background The influenza virus is a highly infectious disease, with a notably rapid transmission rate. Autophagy is triggered by viral infection and is a survival mechanism exerted to maintain cellular homeostasis. Catechin is a representative phenolic acid which exerts anti-inflammatory responses against influenza A virus infection. The aim of this study is to explore the anti-H1N1 influenza virus effects by catechin associated with the restoration of autophagy. Methods XTT assay was used to detect cellular viability. The inhibitory effects on the H1N1 influenza virus were assessed by hemagglutination assay, neuraminidase activity, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The protein levels of H1N1 influenza virulence and autophagic markers were detected by Western blot. Results We herein demonstrated that catechin had no cytotoxic effect on both infected and noninfected A549 cells and exerted protective effects on infected A549 cells. The results of the hemagglutination assay, neuraminidase activity, and qRT-PCR to examine viral load demonstrated that catechin effectively inhibited the replication of the H1N1 influenza virus. The virulent M2 protein and viral nucleoprotein were also inhibited after treatment with catechin. As for the autophagic markers, the LC3B protein was notably decreased by catechin in a dose-dependent manner, while the amount of autophagic vacuoles in H1N1 influenza virus-infected cells also decreased after catechin treatment in a dose-dependent manner. Conclusion Collectively, the autophagy activated by the H1N1 influenza virus could be reversed after catechin treatment. This study indicates that catechin effectively inhibits H1N1 viral proliferation and thus may be applied as an adjuvant in future clinical application.

Pollen has been defined as dietary supplement used to supplement the diet in many countries, but the primary structure and activity of Camellia japonica pollen polysaccharide remain unclear. In this study, the water-soluble polysaccharide extracted from Camellia japonica pollen (WCPP) was fractionated into one neutral fraction (WCPP-N) and two acidic fractions (WCPP-A1 and WCPP-A2) by DEAE-cellulose column, and WCPP-A2 was further fractionated into two homogeneous sub-fractions (WCPP-A2a and WCPP-A2b) by Sepharose CL-6B column. Monosaccharide composition results showed that WCPP-N might mainly contain starch-like glucan as well as some arabinogalactan, while WCPP-A1, WCPP-A2 and its sub-fractions might mainly composed of rhamnogalacturonan I (RG-I) pectic polysaccharide domain backbone with some different types of side chains, including arabinan, galactan, and/or arabinogalactan. The primary structure analysis of WCPP-A2a by NMR spectra analysis suggested that WCPP-A2a was an RG-I-like pectic polysaccharide, branched at the O-4 of Rha residues in the backbone, with α-(1 → 3,5)-L-arabinan as well as type-II arabinogalactan side chain to which were attached. The results of galectin-3-mediated hemagglutination assay indicated that WCPP-A2a exhibited the strongest inhibitory effect on galectin-3 with MIC value around 0.27 μg/mL. These results suggested the potential use of Camellia japonica pollen polysaccharide as a galectin3 inhibitor in functional foods.

Lectins are proteins of non-immunological origin that may play several biological applications, of which we can highlight the anti-inflammatory and antinociceptive activities. In this work, we evaluated the possible effect of orofacial antinociceptive activity of three plant lectins, Dioclea violacea (DVL - Man/Glc-binding), Vatairea macrocarpa (VML - Gal-binding) and PPL (Parkia platycephala - Man/Glc-binding) in adult zebrafish. Acute nociception was induced by menthol (1.2 μM), or capsaicin (4.93 μM) applied into in the upper lip (5.0 μL) of adult wild zebrafish. Zebrafish were pretreated by intraperitoneal injection (20 μL) with vehicle (Control) or lectins (0.025; 0.05 or 0.1 mg/mL) 30 min before induction. The effect of lectins on zebrafish locomotor behavior was evaluated with the open field test. Naive groups (n = 8) were included in all tests. Our results indicate that only PPL presented antinociceptive induced by capsaicin, suggesting the potential clinical application of PPL as inhibitor of orofacial nociception and that this effect may be due to the modulation of TRPV1 channel. In conclusion, lectins that exhibit affinity to the same or different carbohydrates do not necessarily have an antinociceptive effect on the orofacial nociception model, indicating that the glycan carbohydrate binding pattern may be related to the effect on nociception inhibition.

Background This study was performed to investigate the necessity of the second treponemal test and to evaluate the diagnostic performance of the first treponemal test in the reverse algorithm of syphilis diagnosis. Methods Abbott Architect Syphilis TP assay, a chemiluminescence immunoassay (CIA), was used as the first step treponemal test. Treponema pallidum haemagglutination assay (TPHA) test results of reactive samples from the first test were recorded. TPHA test result was considered as confirmatory. TPHA test results were grouped according to their Abbott Architect Syphilis TP assay results and they were compared with Mann-Whitney U test. For Abbott Architect Syphilis TP assay, a cutoff value with 100% specificity was determined via a ROC curve analysis which would render TPHA test unnecessary. Results Out of 146,800 samples 2,646 were reactive in the first step. Of those, 2,002 had a TPHA test result. Of the 2,002 TPHA tests, 1,706 were positive and 296 were negative. TPHA positive and negative groups have significantly different CIA signal/cutoff values. Using a ROC curve built for evaluation of the first-step test, the maximum Youden's index value was found as 5.26. If this value would be accepted as cutoff, it would have a specificity of 85%. The specificity of 100% can be reached if a new cutoff value is set to 27.83. Conclusions Calculated cutoff value with 100% specificity is not practically applicable. It achieves saving of TPHA test in only six percent of reactive samples. Architect Syphilis TP assay is advantageous in large laboratories but is not enough to lead diagnosis without a second treponemal assay. Therefore it was decided to continue the reverse algorithm with dual treponemal assays.

BACKGROUND Red blood cell (RBC) alloantibody titration is a quasi-quantitative method to assess antibody concentration and is considered a useful means of estimating maternal alloimmunization during pregnancy. Traditionally, titration is performed using conventional tube test (CTT). The gel microcolumn agglutination-based method (GMA) has been proven reliable for many immunohematology tests. Our study compared CTT with GMA of two different, commercially available GMA systems for RBC alloantibody titration. METHODS Serum samples with significant RBC-alloantibodies were evaluated in our study. Each sample was titrated concurrently with CTT, with ID-DiaMed-GmbH, Cressier, Switzerland (GMA1), and with DG Gel Coombs Diagnostic Grifols, Passeig Fluvial, Spain (GMA2). RESULTS One hundred thirty-seven titration tests including 50 anti-D, 25 anti-Kell, 10 anti-E, 9 anti-Jka, 8 anti-c, 5 anti-Cw, 5 anti-Fya, 7 anti-M, 6 anti-Kpa, 3 anti-Lua, 1 anti-e, 3 anti-G, and 2 anti-Cha were performed and evaluated. Samples tested by CTT versus GMA1 and GMA2 generated mostly equal or higher titers by GMAs. The results of both comparisons were in good agreement (W = 0.91, p < 0.0001, and W = 0.92, p < 0.0001, respectively). For all antibody specificities, the mean absolute difference in titers ranged from 1 - 3 for both GMA1 and GMA2 versus CTT. Samples tested by GMA1 vs. GMA2 were in almost perfect agreement (W = 0.95, p < 0.0001). CONCLUSIONS Although both GMAs were found slightly more sensitive than CTT for alloantibody titration, the differences were not significant and the agreement between all methods was very good, possibly indicating GMA as a suitable alternative to CTT in RBC antibody titration.

A previously healthy 35-year-old man suffering from behavioral and mental deterioration for three months was referred to our facility. On admission, the patient was dysphasic and had tonic-clonic seizures. Neurological examination showed mental confusion, euphoric mood, mania, paranoia, and mild motor dysphasia. Magnetic resonance imaging (MRI) was performed twice but no abnormalities were revealed. His acute confusional state, elevated cerebrospinal fluid (CSF) protein, epileptic seizure and electroencephalogram showing intermittent frontal slowness were all suggestive of encephalitis. The patient was treated with acyclovir without significant improvement in his condition. Testing for herpes simplex virus and human immunodeficiency virus was negative. Limbic encephalitis was suspected and the patient was treated with intravenous immunoglobulin (IVIG) for five days. Venereal disease research laboratory and Treponema pallidum hemagglutination assay were both tested positive in serum and CSF. Neurosyphilis was diagnosed, and the patient received a 14-day course of penicillin G with gradual improvement. Anti-N-methyl-D-aspartate-receptor (anti-NMDAR) antibodies were positive in serum, confirming the presence of encephalitis. The atypical clinical presentation of neurosyphilis with symptomatology mimicking encephalitis and no MRI abnormalities made the diagnosis challenging. Coexistence of neurosyphilis with anti-NMDAR encephalitis has been reported only in one recent study. Our case demonstrates the importance of testing for syphilis in patients with unexplained neurologic deficits and suspected encephalitis.

A(H1N1) strains of Influenzavirus were responsible for 2 pandemics in the last 100 years. Because infections experienced early in life may have a long-lasting influence on future immune response against other influenza strains, we drew on previously collected seroincidence data from Singapore (n = 2,554; June–October 2009) to investigate whether the 1918 pandemic influenza virus and its early descendants produced an age-related signature in immune responses against the A/California/7/2009(H1N1)pdm09 virus of 2009. Hemagglutination inhibition assays revealed a J-shaped relationship; the oldest birth cohort (born in 1911–1926) had the highest titers, followed by the youngest (born in 1987–1992). Differential response by vaccination history was also observed, with seasonal influenza vaccine being associated with higher titers mainly in the oldest birth cohort. On the assumption that antibody titers are a correlate of protection, structural equation modeling predicted that a titer-mediated effect by the vaccine could, on its own, account for a negative association with seroconversion equivalent to a risk reduction of 23% (relative risk = 0.77, 95% confidence interval: 0.60, 0.99) in the oldest birth cohort. A subset of 503 samples tested against the A/Brisbane/59/2007(H1N1) and A/Puerto Rico/8/1934(H1N1) strains also revealed different age-related antibody profiles. The effectiveness of seasonal influenza vaccines against future pandemic strains could thus be age-dependent and related to early-life exposures.

Portable, on-site blood typing methods will help provide life-saving blood transfusions to patients during an emergency or natural calamity, such as significant earthquakes. We have previously developed waveguide-mode (WM) sensors for forward ABO and Rh(D) blood typing and detection of antibodies against hepatitis B virus and hepatitis C virus. In this study, we evaluated a WM-sensor for reverse ABO blood typing. Since reverse ABO blood typing is a method for detection of antibodies against type A and type B oligosaccharide antigens on the surface of red blood cells (RBCs), we fixed a synthetic type A or type B trisaccharide antigen on the sensor chip of the WM sensor. We obtained significant changes in the reflectance spectra from a WM sensor on type A antigen with type B plasma and type O plasma and on type B antigen with type A plasma and type O plasma, and no spectrum changes on type A antigen or type B antigen with type AB plasma. Signal enhancement with the addition of a peroxidase reaction failed to increase the sensitivity for detection on oligosaccharide chips. By utilizing hemagglutination detection using regent type A and type B RBCs, we successfully determined reverse ABO blood groups with higher sensitivity compared to a method using oligosaccharide antigens. Thus, functionality of a portable device utilizing a WM sensor can be expanded to include reverse ABO blood typing and, in combination with forward ABO typing and antivirus antibody detection, may be useful for on-site blood testing in emergency settings.

Influenza D virus (IDV) has been identified in domestic cattle, swine, camelid, and small ruminant populations across North America, Europe, Asia, South America, and Africa. Our study investigated seroprevalence and transmissibility of IDV in feral swine. During 2012-2013, we evaluated feral swine populations in 4 US states; of 256 swine tested, 57 (19.1%) were IDV seropositive. Among 96 archived influenza A virus-seropositive feral swine samples collected from 16 US states during 2010-2013, 41 (42.7%) were IDV seropositive. Infection studies demonstrated that IDV-inoculated feral swine shed virus 3-5 days postinoculation and seroconverted at 21 days postinoculation; 50% of in-contact naive feral swine shed virus, seroconverted, or both. Immunohistochemical staining showed viral antigen within epithelial cells of the respiratory tract, including trachea, soft palate, and lungs. Our findings suggest that feral swine might serve an important role in the ecology of IDV.

Background: Screening for Trypanosoma cruzi infection was performed amongst children in a rural community in the Bolivian Chaco, an area known for high prevalence. The force of infection (FOI) was estimated. Methods: A total of 423 children attending the local school were screened using the Chagas Detect Plus (CDP) rapid test (InBios International, Inc.). CDP-positive specimens were further tested by indirect haemagglutination assay (IHA) and Wiener Recombinante v3.0 ELISA. A catalytic model was used to estimate FOI. Results: Confirmed seroprevalence was 0.22, rising steeply with age. The mean age of seropositive individuals was 13 years. The calculated specificity of the rapid test was 91.9%. The annual incidence estimated from the FOI was 0.021. Conclusions: This study demonstrates persistent transmission and continuing high levels of T. cruzi infection in the Bolivian Chaco, and highlights the practicality of school-based screening. Keywords: Chagas disease, Trypanosoma cruzi, T. cruzi, Bolivian Chaco, Screening study

Hydatic cyst is a crucial and prevalent parasitic disease in the developing countries in the Mediterranean region. Its diagnosis is sometimes problematic because of non-specific complaints and unavailability of any positive results in a routine laboratory analysis. Isolated renal hydatid cysts are very rare. In this study, a primary left renal hydatid cyst which was found in a 12-year-old boy was presented. He was referred by another hospital to our department with a flank pain and cystic mass in left kidney. Indirect haemagglutination test (IHA) for Echinococcus was negative. Nephrectomy was performed with the diagnosis of renal cyst hydatic. Renal cyst hydatid may present with various clinical findings ranging from asymptomatic clinical course to total loss in renal function. It will be beneficial to consider a renal hydatid cyst in patients with blurred flank pain, even if IHA is negative.

Hydatid disease is a parasitic disease. Although the disease causes many organ involvement, intracranial involvement is rarely seen. Clinical findings vary according to the number, size, localization of the cyst and the immune status of the patient. We aimed to present a rare case with intracranial hydatid cyst. A 8-year-old male patient presented with headache and vomiting. Physical examination revealed papillary edema. Brain computerized tomography imaging revealed a 13x13 cm cystic formation in the brain. The patient’s echinococcal indirect hemagglutination test result was positive. The patient was started on oral albendazole therapy for intracranial hydatid cyst and was operated by brain surgery. Pathologic examination of the cyst was compatible with hydatid cyst. Although cysts are detected in the liver and lungs, cysts may be seen in atypically located areas. It should be kept in mind in clinically suspected patients that the disease can develop in rare organs, especially in patients with endemic areas. In patients with hydatid disease, advanced radiological examinations such as lung X-ray, abdominal ultrasonography and brain magnetic resonance imaging and echocardiographic examination should be performed for other organ involvement.

Methicillin resistant Staphylococcus aureus (MRSA) is a versatile pathogen capable of causing multitude of human diseases. It is one of the most important nosocomial pathogen that implicated in community and healthcare associated infections. Therefore, this study aims to characterize different SCCmec elements found in MRSA isolates. Moreover, molecular typing was performed to investigate the genetic relatedness among MRSA isolates.Phenotypic identification of MRSA was done by disc diffusion method. The MRSA isolates were typed based on the SCCmec, coa and agr genes. Phenotypic characterization included the detection of biofilm, lipase, protease, lecithinase, staphylokinase and hemagglutination. Also, hla, hlb, hlg, hld, tsst-1, psm-mec and mecI genes were detected genotypically. The correlation between the molecular types identified and the profile of virulence factors, clinical and geographical sources was determined for all isolates.Eighty five isolates were identified as MRSA. Eight types of SCCmec elements were detected among these isolates. Type V was the most observed type (56.47%). Regarding the correlation between SCCmec types and virulence factors, type V SCCmec exhibited a significant association with biofilm (p 0.0001), staphylokinase (p = 0.0495) and tsst-1 (p = 0.0498). Molecular typing of coa gave an insight to the presence of specific types in specific hospital wards. Based on agr typing, agr I was the highest prevalent type in MRSA isolates (54.11%).There is an increase of MRSA infections particularly the community acquired with high variability in the distribution of virulence factors among different SCCmec types. The association between type III and V SCCmec with certain hospitals may be an evidence of nosocomial infection among these hospitals.

Syphilis prevalence has been declining in every region by several percentage points per year. Nevertheless, the prevalence by region continued to vary, from as little as 0.1% in the European Region to as high as 3% in the African Region., Background This study assessed levels, trends, and associations of observed syphilis prevalence in the general adult population using global pooled analyses. Methods A standardized database of syphilis prevalence was compiled by pooling systematically gathered data. Random-effects meta-analyses and meta-regressions were conducted using data from the period 1990–2016 to estimate pooled measures and assess predictors and trends. Countries were classified by World Health Organization region. Sensitivity analyses were conducted. Results The database included 1103 prevalence measures from 136 million syphilis tests across 154 countries (85% from women in antenatal care). Global pooled mean prevalence (weighted by region population size) was 1.11% (95% confidence interval [CI], .99–1.22). Prevalence predictors were region, diagnostic assay, sample size, and calendar year interacting with region. Compared to the African Region, the adjusted odds ratio (AOR) was 0.42 (95% CI, .33–.54) for the Region of the Americas, 0.13 (95% CI, .09–.19) for the Eastern Mediterranean Region, 0.05 (95% CI, .03–.07) for the European Region, 0.21 (95% CI, .16–.28) for the South-East Asia Region, and 0.41 (95% CI, .32–.53) for the Western Pacific Region. Treponema pallidum hemagglutination assay (TPHA) only or rapid plasma reagin (RPR) only, compared with dual RPR/TPHA diagnosis, produced higher prevalence (AOR >1.26), as did smaller sample-size studies (2.16). Prevalence declined in all regions; the annual AORs ranged from 0.84 (95% CI, .79–.90) in the Eastern Mediterranean to 0.97 (95% CI, .97–1.01) in the Western Pacific. The pooled mean male-to-female prevalence ratio was 1.00 (95% CI, .89–1.13). Sensitivity analyses confirmed robustness of results. Conclusions Syphilis prevalence has declined globally over the past 3 decades. Large differences in prevalence persist among regions, with the African Region consistently the most affected.

John Cunningham virus (JCPyV) is an ubiquitous human pathogen that causes disease in immunocompromised patients. The JCPyV genome is composed of an early region and a late region, which are physically separated by the non-coding control region (NCCR). The DNA sequence of the NCCR distinguishes two forms of JCPyV, the designated archetype and the prototype, which resulted from a rearrangement of the archetype sequence. To date, the cell culture systems for propagating JCPyV archetype have been very limited in their availability and robustness. Prior to this study, it was demonstrated that JCPyV archetype DNA replicates in COS-7 simian kidney cells expressing SV40 TAg and COS-7 cells expressing HIV-1 Tat. Based on these observations, the present study was conducted to reproduce an in vitro model in COS-7 cells transfected with the JCPyV archetype strain in order to study JCPyV DNA replication and analyze NCCR rearrangements during the viral life cycle. The efficiency of JCPyV replication was evaluated by quantitative PCR (Q-PCR) and by hemagglutination (HA) assay after transfection. In parallel, sequence analysis of JCPyV NCCR was performed. JCPyV efficiently replicated in kidney-derived COS-7 cells, as demonstrated by a progressive increase in viral load and virion particle production after transfection. The archetypal structure of NCCR was maintained during the viral cycle, but two characteristic point mutations were detected 28 days after transfection. This model is a useful tool for analyzing NCCR rearrangements during in vitro replication in cells that are sites of viral persistence, such as tubular epithelial cells of the kidney.

Crude soluble antigen (CSA) produced from Entamoeba histolytica trophozoite is conventionally used for serodiagnosis of invasive amoebiasis. However, high background seropositivities by CSA-assay in endemic areas complicate the interpretation of positive result in clinical settings. Instead, incorporating a second assay which indicates active or recent infection into the routine amoebic serology could possibly complement the limitations of CSA-assay. Hence, the present study aimed to evaluate the diagnostic efficacies of indirect ELISAs using CSA and excretory-secretory antigen (ESA) for serodiagnosis of amoebic liver abscess (ALA). Reference standard for diagnosis of ALA at Hospital Universiti Sains Malaysia is based on clinical presentation, radiological imaging and positive indirect haemagglutination assay (titer ≥256). Five groups of human serum samples collected from the hospital included Group I - ALA diagnosed by the reference standard and pus aspirate analysis using real-time PCR (n=10), Group II - ALA diagnosed by the reference standard only (n=41), Group III - healthy control (n=45), Group IV - other diseases control (n=51) and Group V - other infectious diseases control (n=31). For serodiagnosis of ALA serum samples (Group I and II), CSA-ELISA showed sensitivities of 100% for both groups, while ESA-ELISA showed sensitivities of 100% and 88%, respectively. For serodiagnosis of non-ALA serum samples (Group III, IV and V), CSA-ELISA showed specificities of 91%, 75% and 100%, respectively; while ESA-ELISA showed specificities of 96%, 98% and 100%, respectively. Indirect ELISAs using CSA and ESA have shown distinct strength for serodiagnosis of ALA, in terms of sensitivity and specificity, respectively. In conclusion, parallel analysis by both assays improved the overall efficacies of amoebic serology as compared to either single assay.

Objective In this study, forming of experimental toxoplasmosis in quails; clinical, pathological, and serological determination of tissue lesions and bioassay techniques, which were aimed to compare them and determine pathogenesis. Methods A total of 120 one-year-old female quails (Coturnix coturnix japonica) were divided into oral infection, parenteral infection, and control groups. The oral group was infected with 0.5 ml inoculum suspension containing 106 tachyzoites of Toxoplasma gondii, whereas the control group was administered 0.5 ml of saline. The parenteral group was further divided into the following four subgroups: intraperitoneal, intramuscular, intravenous, and cloacal. The quails of the parenteral group were also divided into two groups and one by control group within itself for the 105 and 104 doses of the tachyzoite inoculums. Results Because of acute toxoplasmosis, death occurred in a quail that as intramuscularly infected with 105 tachyzoites; the quail exhibited neurological clinical symptoms such as torticollis, ataxia, and tremor. In histopathologic examination, T. gondii tissue cysts were detected in infected quails that were intramuscularly infected with 105 tachyzoites. Mouse trials were conducted using tissues of seropositive quails and isolated from peritoneal fluids infected mice. By Sabin-Feldman dye test and indirect hemagglutination test, seropositivity was observed in quails infected with 105 and 104 tachyzoites. Conclusion Similar studies and subclinical cases, which may overlooked was concluded for diagnosis of toxoplasmosis with useful bioassay applications and serological tests.

OBJECTIVE Glycine max is commonly used in Algeria for treatment of anemia deficiency and osteoporosis, it ranks first in terms of vegetal proteins. The experiment was aimed at characterizing the proteinaceous Glycine max extract and evaluating its antioxidant, biological and hematological potential. METHODOLOGY Extraction of proteinaceous materials from Glycine max plant was undertaken using water and n-hexane as extracting media. The isolation of proteins from the crude materials was done, providing the use of ammonium sulfate. The Glycine max proteins were characterized by UV-visible and FT-IR spectroscopy and analyzed by SEM micrograph and x-ray diffraction (XRD). Rheological parameters G' and G'' were assessed. The isolated proteins were tested for their antioxidant, antimicrobial and hemagglutination activities. RESULTS There was a gelling effect of the protein extract which can be used as an alternative in principally made vaccines with its microbiological and antifungal activities. CONCLUSION The proteinaceous extract from Algerian Glycine max would have a potential use in biomedical application.

5 páginas, 3 tablas, The diagnosis of cystic echinococcosis (CE) is based on imaging. Serology supports imaging in suspected cases, but no consensus exists on the algorithm to apply when imaging is inconclusive. We performed a retrospective analysis of serology results of patients with untreated hepatic CE and non-CE lesions, seen from 2005 to 017, to evaluate their accuracy in the differential diagnosis of hepatic CE. Serology results of three seroassays for echinococcosis (ELISA RIDASCREEN, indirect hemagglutination (IHA) Cellognost, and Western blot LDBIO) and clinical characteristics of eligible patients were retrieved. Patients were grouped as having active or inactive CE and liquid or solid non-CE lesions. Sensitivity, specificity, and diagnostic accuracy were compared between scenarios encompassing different test combinations. Eligible patients included 104 patients with CE and 257 with non-CE lesions. Sensitivity and diagnostic accuracy ofWestern blot (WB)were significantly higher than those of the following: 1) IHA or ELISA alone, 2) IHA+ELISA interpreted as positive if both or either tests positive, and 3) IHA+ELISA confirmed by WB if discordant. The best performances were obtained when WB was applied on discordant or concordant negative IHA+ELISA. Analyses performed within “active CE (n = 52) versus liquid non-CE (n = 245)” and “inactive CE (n = 52) versus solid non-CE (n = 12)” groups showed similar results. Specificity was high for all tests (0.99–1.00) and did not differ between test combination scenarios. WB may be the best test to apply in a one-test approach. Two first-level tests confirmed by WB seem to provide the best diagnostic accuracy. Further studies should be performed in different settings, especially where lower test specificity is likely., This work was funded by the European Union Seventh Framework Program (FP7/2007–2013) under the project HERACLES, grant agreement no. 602051–to E. B.

We present our findings in two cases of retinal dysfunction caused by syphilitic outer retinopathy. CASE 1: A 59-year-old man visited our clinic complaining of blurred vision in his left eye. Optical coherence tomography (OCT) demonstrated an absence of the ellipsoid zone (EZ) in the left eye. A round yellowish-white lesion was observed in the posterior pole of the left fundus. Fundus autofluorescence (FAF) showed hyperfluorescent areas in the posterior pole of both fundi although no specific ophthalmoscopic findings were seen in the right eye. The amplitudes of the LA 3.0 1 Hz and LA 3.0 30 Hz ERG responses were reduced with better preservation of the rod responses. Based on a strong positivity to the rapid plasma reagin (RPR) assay and the Treponema pallidum hemagglutination (TPHA) test, he was diagnosed with syphilitic outer retinopathy and treated with systemic antibiotics. The treatment resulted in a restoration of the retinal structures and cone function. CASE 2: A 47-year-old man was referred to our clinic complaining of reduced vision in both eyes. Although the ocular fundus appeared normal, FAF showed a diffuse hyperfluorescent area in the posterior pole and multiple hyperfluorescent spots. Indocyanine green angiography showed multiple confluent areas of hypofluorescence. OCT demonstrated irregular EZs in both eyes. The amplitudes of the LA 3.0 1 Hz and LA 3.0 30 Hz ERG responses were slightly reduced with prolonged implicit times. These findings are comparable to the findings in patients with multiple evanescent white dot syndrome. However, the strong positivity to the RPR and TPHA tests led us to diagnose the patient with outer retinopathy caused by syphilis. Systemic administration of antibiotics resulted in the restoration of the retinal structures and retinal function.Syphilitic outer retinopathy affected the retinal structures and function that can be restored by antibiotic treatments.

The separation of plasma from blood cells is critical for the accuracy of blood tests because cellular fractions can create discrepancies in analysis. The most common method to separate blood cells from the liquid part of the blood is centrifugation, which is not always applicable in resource-constrained areas and countries. In this study, we describe the generation of monoclonal antibodies (mAbs) against glycophorin A (GPA) of human erythrocytes. BALB/c mice were immunized with human erythrocytes followed by purified GPA. The splenocytes of the immunized mice were fused with Sp2/0 myeloma cells by hybridoma technique. Hybridoma clones were screened by hemagglutination assay and enzyme-linked immunosorbent assay (ELISA). Six hybridoma clones were obtained and subcloned. The characterization of the purified mAbs demonstrates that they are able to bind and retain erythrocytes in hemagglutination assay. Furthermore, one of the mAbs 1A9 recognizes purified GPA in ELISA, whereas the other mAb 1G7 is able to immunoprecipitate GPA from human erythrocyte lysates, and a band of 38 kDa is detected. In conclusion, the anti-GPA mAbs are useful tools in developing a quick and easy way to separate blood plasma from whole blood for clinical tests, and in developing bi-specific antibodies for other clinical applications.