Serological cross-sectional survey of equine infectious anemia in Saudi Arabia.

The equine infectious anaemia virus (EIAV) is one of the most serious equine diseases worldwide. There is scarce information on the epizootiology of equine infectious anaemia (EIA) in Saudi Arabia. Given the importance of the equine industry in Saudi Arabia, this cross- -sectional study aims to provide information about the prevalence of EIAV based on serological surveillance of the equine population in the country. A total of 4728 sera samples were collected (4523 horses and 205 donkeys) between December 2017 and November 2019. All samples were tested using commercially available EIAV ELISA. All tested samples showed negative results for EIAV antibodies with a 95% confidence interval. The results provided evidence that Saudi Arabia's equine populations (horses and donkeys) are currently free of EIAV. The results also suggest the need for continuous monitoring of EIAV and strict regulation when importing horses from other countries.

Outbreak investigation and molecular diagnosis of Lumpy skin disease among livestock in Saudi Arabia 2016.

Lumpy skin disease (LSD) is a highly infectious disease of cattle caused by a virus of the Capripoxvirus genus in the family Poxviridae. The disease is a major concern for the dairy industry in Saudi Arabia. In this study, an outbreak of LSD in cattle herds was detected in Saudi Arabia in 2016. LSD outbreak was investigated in five regions of Saudi Arabia: Al-Hassa, Al-Sharqia, Al-Qassim, Riyadh and Al-Taif during the period from April to July 2016. Tissues from skin nodules were collected to characterize the virus by a real-time polymerase chain reaction (rt-PCR). During this period, 64,109 cattle were examined and morbidity, mortality and case fatality rates were 6%, 0.99% and 16.6%, respectively. The analysis showed 3,852 infected cases and 641 deaths. highest number of infected animals was reported in Al-Hassa (2,825), followed by Al-Qassim (547), Riyadh (471), Al-Sharqia (6) and Al-Taif (3). The highest morbidity rates were observed in Al-Qassim (6.8%), Al-Hassa (6.2%), Riyadh (5.5%) and Al-Taif (0.96%), while the lowest morbidity rates were recorded in Al-Sharqia (0.27%). The highest mortality rates were also observed in Al-Qassim (2.3%), followed by Al-Hassa (0.97%), Riyadh (0.19%) and lowest in Al-Sharqia and Taif (0%). LSD virus was detected in all samples (n = 191) by real-time PCR analysis. The disease has been observed in the cattle regardless of previous vaccination using the locally Romanian-pox vaccine; therefore, vaccination programme and vaccine efficacy should be assessed under field conditions.

Application of a loop-mediated isothermal amplification (LAMP) assay for molecular identification of Trueperella pyogenes isolated from various origins.

In the present study 28 Trueperella pyogenes strains isolated from various origins could successfully be identified with a newly designed loop-mediated isothermal amplification (LAMP) assay based on gene cpn60 encoding chaperonin. No cross reaction could be observed with control strains representing four species of genus Trueperella and seven species of closely related genus Arcanobacterium. The present cpn60 LAMP assay might allow a reliable and low cost identification of T. pyogenes also in laboratories with less specified equipment.

Cervical Ectopic Pregnancy: A Management Dilemma

A 28-year-old G3P1+1 at 6 weeks period of amenorrhea with a previous Caesarean section presented with per vaginal bleeding. A cervical ectopic pregnancy was confirmed by a transvaginal scan. An intramuscular methotrexate was given followed by intracervical route due to poor decline of the serum βHCG. However, due to persistent increment of serum βHCG, an additional four doses of intramuscular methotrexate with folinic acid rescue were administered and she responded well to the regime. Unfortunately, following the last dose, she developed an episode of excessive per vaginal bleeding which required suction and curettage of the cervical canal. A Foley‘s catheter balloon was placed intracervically as a tamponade and the bleeding was successfully arrested.

Uterine Sex Cord Tumour- Management Dilemma

Uterine sex cord tumour is a very rare tumour with uncertain management strategies and prognosis. A 61-year-old, nulliparous, who was not on hormone replacement therapy, presented with first episode of postmenopausal bleeding. A transvaginal scan revealed an enlarged uterus with thick endometrial lining and features of multiple degenerated fibroid. Endometrial biopsy was negative for malignancy. Computed tomography of the abdomen and pelvis confirmed the mass, with atrophic ovaries and incidental finding of bilateral hydronephrosis requiring stentings. Otherwise, there were no pelvic lymph nodes enlargement. Our impression was a uterine sarcoma and we decided for total abdominal hysterectomy with bilateral salpingooophorectomy. Surprisingly, the histology report confirmed uterine sex cord tumour. There are less cases of recurrence and there is no general consensus on the management. However, we decided for adjuvant chemotherapy (BEP regime) as the malignant cells infiltrated more than half of myometrial thickness, with good outcome.

Case Review: Post Partum Vulvo-vaginal Haematomas and Options of Management

Vulvo-vaginal haematomas are not an uncommon obstetric complication. Despite advances in obstetric care, practice and technique, vulvo-vaginal haematomas do occur especially in complicated vaginal deliveries. Various management options are available for vulvo-vaginal haematomas. We describe three cases of vulvo-vaginal haematomas with different severity and presentations which were managed in different manners i.e. local haemostasis control, laparotomy with hysterectomy, and transarterial embolisation. The choice of treatment options would mainly depend on the clinical presentations, availability of expertise as well as facilities. Early identification is crucial.

Unruptured Left Cornual Pregnancy with a High Serum β-hCG: A Case Report

Ectopic pregnancy in the interstitial part of the fallopian tube (cornual pregnancy) is a rare condition but can be fatal. Traditionally, the treatment had been cornual resection or hysterectomy. More conservative approaches had been advocated recently. There is no consensus on the dose or number of methotrexate injections that should be used in the treatment of interstitial pregnancies. Single dose intramuscular methotrexate is one of the treatment options. However, the failure rate is higher if the serum β-hCG (beta-human chorionic gonadotrophin) level is more than 5000 IU/L. We report a case of cornual ectopic pregnancy with high initial serum β-hCG level being successfully treated with multiple doses of systemic methotrexate. MRI was used to assess clinical resolution of cornual ectopic pregnancy.

Morbidly Adherent Placenta at Extreme Prematurity: Can Major Haemorrhage and Hysterectomy be Prevented?

Morbidly adherent placenta with spontaneous rupture of membrane at extreme prematurity poses poor pregnancy outcome. Various issues on different management modalities still remain perplexed and individual consideration is vital. Two cases of morbidly adherent placenta with symptomatic per vaginal bleeding and spontaneous rupture of membrane at severe prematurity were reviewed and discussed. We found that, active intervention by termination of pregnancy and methotrexate therapy at early gestation can prevent the need of hysterectomy following major obstetrics haemorrhage.

Neuraminidase treatment abrogates the binding abnormality of IgA1 from IgA nephropathy patients and the differential charge distribution of its alpha-heavy chains.

We have studied the interaction of the Gal-GalNAc-reactive champedak lectin-C with neuraminidase-treated and untreated IgA1 from IgA nephropathy patients. The binding ability of the lectin to untreated IgA1 from IgA nephropathy patients was significantly lower as compared to the untreated IgA1 from normal controls. This differential lectin-binding capacity was abrogated when the experiment was performed on neuraminidase-treated sera. Treatment of the serum IgA1 with neuraminidase also abrogated the differential charge distribution between the alpha-heavy chains of IgA nephropathy patients and normal controls.

The application of Artocarpus integer seed lectin-M in the detection and isolation of selective human serum acute-phase proteins and immunoglobulins.

Champedak (Artocarpus integer) lectin-M is a lectin with high specificity and affinity for the core-mannosyl residues of the N-linked oligosaccharides of glycoproteins. We have studied the interaction of the champedak seed lectin with human serum glycoproteins that were resolved by 2-dimensional (2-D) gel electrophoresis. The lectin demonstrated strong interaction with haptoglobin beta chain, orosomucoid, alpha1-antitrypsin, alpha2-HS glycoprotein, transferrin, hemopexin, alpha1B-glycoprotein, and the heavy chains of IgA, IgM and IgG of the human serum. With exceptions of the heavy chains of the immunoglobulins and alpha1B-glycoprotein, all the other lectin-M-probed glycopeptides are acute-phase proteins. The use of champedak lectin-M to probe for serum glycoproteins that were separated in a 2-D gel electrophoresis and Western blotting technique may be conveniently applied to analyse the acute-phase and humoral immune responses simultaneously. Subjecting human serum to immobilised-lectin-M affinity chromatography was able to isolate intact haptoglobin, alpha1-antitrypsin, alpha1B-glycoprotein, hemopexin and IgA.

The interaction of selective plant lectins with neuraminidase-treated and untreated IgA1 from the sera of IgA nephropathy patients.

A study on the binding interaction of lectins from Artocarpus heterophyllus (jacalin), Glycine max and Sambucus nigra with standardised quantity of IgA from the IgA nephropathy patients and normal controls was performed. The Glycine max lectin demonstrated higher affinity towards the serum IgA of IgAN patients as compared to normal controls. However, the affinity binding was lower in cases ofjacalin and the Sambucus nigra lectin. When serum samples were treated with neuraminidase, the differential jacalin affinity binding between IgA1 of patients and normal controls was abrogated. Our data are in support of the view that the O-linked oligosaccharide moieties of the patients IgA1 were generally lacking in galactose and sialic acid residues.

Effect of the mannose-binding Artocarpus integer lectin on the cellular proliferation of murine lymphocytes.

The effect of the mannose-binding champedak (Artocarpus integer) lectin-M on the cellular proliferation of murine lymphocytes was investigated in this study. Our data demonstrated that the lectin was the main mitogenic component in the crude extract of the champedak seeds. It stimulated the proliferation of murine T cells at an optimal concentration of 2.5 microg/ml in a 3 day culture. Lectin-M appeared to be a T-cell mitogen as it does not induce significant DNA synthesis when cultured with spleen cells from the nude mouse. In the absence of T cells, the lectin was incapable of inducing resting B cells to differentiate into immunoglobulin secreting plasma cells.

Sera of IgA nephropathy patients contain a heterogeneous population of relatively cationic alpha-heavy chains.

Sera of IgA nephropathy (IgAN) patients and normal subjects were analysed by two-dimensional (2-D) gel electrophoresis. Densitometric analysis of the 2-D gels of IgAN patients and normal subjects revealed that their protein maps were comparable. There was no shift of pI values in the major alpha-heavy chain spots. However, the volume of the alpha-heavy chain bands were differently distributed. Distribution was significantly lower at the anionic region in IgAN patients (mean anionic:cationic ratio of 1.184 +/- 0.311) as compared to normal healthy controls (mean anionic:cationic ratio of 2.139 +/- 0.538). Our data are in support of the previously reported findings that IgA1 of IgAN patients were lacking in sialic acid residues.

Isolation of a mannose-binding and IgE- and IgM-reactive lectin from the seeds of Artocarpus integer.

A mannose-binding lectin, termed champedak lectin-M, was isolated from an extract of the crude seeds of champedak (Artocarpus integer). On gel filtration chromatography, the lectin eluted in a single peak at elution volumes corresponding to 64 kDa. SDS-PAGE showed the mannose-binding lectin to be composed of 16.8 kDa polypeptides with some of the polypeptides being disulphide-linked to give dimers. When tested with all isotypes of immunoglobulins, champedak lectin-M demonstrated a selective strong interaction with human IgE and IgM, and a weak interaction with IgA2. The binding interactions of lectin-M were metal ion independent. The lectin was also shown to interact with horseradish peroxidase, ovalbumin, porcine thyroglobulin, human alpha1-acid glycoprotein, transferrin and alpha1-antitrypsin. It demonstrated a binding preference to Man alpha 1-3Man ligands in comparison to Man alpha 1-6Man or Man alpha 1-2Man.

Effect of Artocarpus integer lectin on functional activity of guinea-pig complement.

The effect of Artocarpus integer lectin (lectin C) on the functional activity of guinea-pig complement was investigated. Purified and crude extract of lectin C from six cultivars of Artocarpus integer seeds were found to consume complement and thus decreased the complement-induced haemolytic activity of sensitized sheep erythrocytes. The change in the complement-mediated haemolytic activity was significantly decreased when incubation of the lectins was performed in the presence of melibiose. The reversal effect of the carbohydrate, which is a potent inhibitor of the lectin's binding to O-linked oligosaccharides of glycoprotein, demonstrate involvement of the lectins interaction with O-glycans of glycoproteins in the consumption of guinea-pig complement.

Comparative analyses of IgA1 binding lectins from seeds of six distinct clones of Artocarpus integer.

Purified lectins from seeds of six distinct clones of Artocarpus integer (lectin C) were shown to be structurally and functionally similar. All lectins comprised of two types of non-covalently-linked subunits with apparent M(r) of 13,300 and 16,000. The lectins appeared to interact with several human serum proteins, with the predominance of the IgA1 and C1 inhibitor molecules. Interaction was not detected with IgA2, IgD, IgG and IgM. The lectin Cs were also shown to precipitate monkey, sheep, rabbit, cat, hamster, rat and guinea-pig serum. Due to their uniform properties, lectin C may provide better alternative to the Artocarpus heterophyllus lectin, jacalin, for use in future investigations.

Interaction of Artocarpus lectins with human IgA does not involve asparagine-linked oligosaccharide of the immunoglobulin.

In view of the controversy with respect to the interaction of jacalin with human IgA2, a study was undertaken to assess the reactivity of the Artocarpus heterophyllus lectin, as well as the lectin from Artocarpus integer (lectin C), with subclasses of human immunoglobulin A by ELISA. Our data is consistent with the view that Artocarpus lectins have no affinity for the IgA2 immunoglobulins. In further support of the findings, we have established that N-linked oligosaccharide moieties of IgA have no significant bearing in the lectin-immunoglobulin binding. Interaction was also not affected in the presence of 1% (w/v) BSA.

Lectin extracts of champedak seeds demonstrate selective stimulation of T lymphocyte proliferation.

The effect of extracts of champedak (Artocarpus integer) seed lectin on the proliferation of normal human lymphocyte was investigated. The IgA1 binding lectin was demonstrated to stimulate the proliferation of human peripheral blood mononuclear cells. Action of the lectin on enriched T and B cell populations demonstrated T lymphocyte specificity. The lectin was not mitogenic to B lymphocytes. Optimal stimulation of proliferative response was achieved when cells were subjected to 5 days exposure to the crude lectin at 20 micrograms/ml.

Actions of three clostridial IgA proteases on distinct forms of immunoglobulin A molecules.

Three bacterial species of Clostridium (septicum, tertium and sporogenes) were identified to produce extracellular proteases cleaving IgA to Fab and Fc fragments, as demonstrated by SDS-PAGE and immunoelectrophoretic procedures. These enzymes acted on monometric IgA1 paraproteins and normal serum IgA1 but had no activity on IgA2 paraproteins and intact secretory IgA1 from human colostrum. Their action on polyclonal serum IgA1 suggested the absence of neutralizing anti-clostridial IgA protease activity. Although the enzymes were shown not to act on secretory IgA1, they were, however, able to digest free alpha-heavy chains of the dimeric IgA molecules. Susceptibility of the alpha-heavy chain to the proteases was more likely due to the change to a more accessible conformation than because of the absence of neutralizing anti-enzymic activity.