Immunochemical and Biochemical Analysis of Larval Secreted Antigens from the Parasitic Nematodes Ascaris suum and Ascaris lumbricoides

There is a relatively small amount of experimental data regarding the in vitro - released secretory products (IVRS) of the parasitic roundworm nematodes of man and swine, Ascaris lumbricoides and Ascaris suum respectively. The importance of detailed biological examination of the IVRS from these nematodes is justified by the relative inadequacy, for serodiagnosis or prophylaxis, of alternative sources of nematode-derived material: for instance, cuticular surface molecules, or worm homogenates. Here the IVRS from L2 and L3/4 stages of Ascaris lumbricoides and Ascaris suum were examined for the first time both immunochemically and biochemically to evaluate their usefulness for specific detection of ascariasis and for possible use in vaccination of animals or humans against Ascaris infection. The main techniques used involved the extrinsic (125I) or intrinsic (35[S]-methionine) labelling of parasite secretions followed by precipitations with either lectins or immune sera and final analysis of the precipitated nematode products by SDS-PAGE. Alternatively, radio-iodinated IVRS from A. suum were incubated with various proteinases and the products of the reactions also analysed by SDS-PAGE. The first studies of radio-iodinated A. suum IVRS showed that the range and number of worm products were different as the parasites developed from the L2 stage to lung stages (L3/4) and that there was considerable, but incomplete stage-specificity of the repertoire of secretions produced. S. aureus-mediated radio-immunnoprecipitations (SRIP's) of A. suum-immunised rabbit antisera and A. suum IVRS showed that, except for one product from A. suum L3/4 IVRS, all the secretions of A. suum L2 and L3/4 worms are antigenic. The non-antigenic component of L3/4 products was identified as rabbit serum albumin. Antisera from A. suum-immunised mice and rats also precipitated components of larval IVRS in SRIP's but different distinct sets of antigens from those recognized by infected rabbits. One possible reason for these differences could be attributed to the separate genetic composition of each species. Mouse anti-A. suum antisera failed to immunoprecipitate an Mr 14000 component of A. suum larval secretions, which was also found to be the major antigen of adult A. suum body fluid (ABF). As ABF is a commonly used source of material for immunological analysis of Ascaris infection, it could be advisable to look for alternative sources of target antigens that avoid an undue bias towards the Mr 14000 product. Immunological crossreactions have been a major stumbling block for specific detection of nematode parasite infection of humans and animals. One of the ways in which crossreactions could lead to false positive results in serodiagnostic immunoassays is the possible occurence of similar antigens among the products of different species of parasites. The secreted products of A. suum and A. lumbricoides were examined by radio-iodination followed by SDS-PAGE. The results showed that the secretions of larval stages of both species were very similar in SDS-PAGE profile except that IVRS from the larvae of the human roundworm apparently contained a secretion of Mr 17000 that was unique to these. SRIP's with sera from rabbits infected either by A. suum or A. lumbricoides and 125I-labelled IVRS of the larval stages of these parasites followed by examination of the immunoprecipitates by SDS-PAGE, showed that there was complete crossreactivity of a given antiserum of one species and the antigens of the other, including the Mr 17000 antigen of A. lumbricoides. Additionally there was also extensive crossreactivity of antiserum from rabbits infected with the ascaridoidean parasite Toxocara canis and the secreted antigens of both A. suum and A. lumbricoides. These findings advised against the use of IVRS from Ascaris species in possible serodiagnostic assays or in serological tests to differentiate between A. suum and A. lumbricoides as separate species. However it might be possible, by raising monoclonal antibodies (McAb) against Ascaris IVRS to find McAb's that differentiate between the Ascaris species and/or provide specificity in serodiagnostic assays. Radio-iodination of IVRS from Ascaris species involved material collected from the supernatants of worm cultures in vitro, not necessarily representative of the products of living worms, but could have consisted of worm somatic components. One biochemical method that aid in confirming that, for the most part, living worms synthesised the IVRS is the technique of intrinsic (or metabolic) labelling of Ascaris larval products. (Abstract shortened by ProQuest.)

Determinants of anxiety and depression among university teachers during third wave of COVID-19

Abstract Background To estimate the determinants of anxiety and depression among university teachers in Lahore, Pakistan, during COVID-19. Methods A cross-sectional study was conducted by enrolling 668 teachers from the universities of Lahore, Pakistan. Data were collected using a questionnaire. Chi-square for significance and logistic regression for the association were used. Results Majorly, the university teachers, with an average age of 35.29 years, had regular jobs (72.8%), job experience of > 6 years (51.2%) and good self-reported health (55.4%). The majority of the teachers were working as lecturers (59.6%), lecturing in arts (33.5%) or general science (42.5%) departments, having MPhil (37.9%) or master (28.9%) degrees, and teaching via synchronous video (59.3%) mode. Anxiety and depression, severe and extremely severe, were higher among lecturers, MPhil or master degree holders, teachers lecturing arts and general science subjects, and in those on contract employment. Anxiety was significantly associated with academic departments; arts (OR;2.5, p = 0.001) and general science (OR;2.9, p = 0.001), poor health status (OR;4.4, p = 0.018), and contractual employment (OR;1.8, p = 0.003). Depression was associated with academic departments; arts (OR;2.7, p = 0.001) and general science (OR;2.5, p = 0.001), and health status (OR;2.3, p = 0.001). Conclusion Among university teachers, anxiety and depression, severe and extremely severe, were prevalent among lecturers having MPhil or master degrees, belonging to arts and general science departments, and among contract employees. Anxiety and depression were significantly associated with academic disciplines, lower cadre, and poor health status

Relative potential of rhizobium species to enhance the growth and yield attributes of cotton (Gossypium hirsutum L.)

Legumes compensate mineral fertilizer by fixing nitrogen due to the specialized structures i.e. nodules by Rhizobium species. Literature revealed that legumes fixed nitrogen due to Rhizobium inoculation from 50-300 kg NPK ha-1 year-1. Rhizobium besides nitrogen fixation, solubilized phosphates, produced growth hormones and due to its root colonizing ability improved the growth and yield of non-legumes also and performed as plant growth promoting rhizobacteria (PGPR). Study was conducted to assess the relative efficiency of Rhizobium species for the growth and yield of cotton. Different isolates of five species of Rhizobium species responsible for different nodule formation in legumes were assessed for the auxin biosynthesis potential as IAA equivalents and isolates having higher values for IAA equivalents were used for experimentation. Results revealed that isolates of Rhizobium species improved the growth and physiological parameters of cotton. Higher values were root/shoot length and mass were observed with Rhizobium species of berseem (Br5). Bacterial inoculation with isolate (Br5) produced 60.94, 64.40 g shoot/root mass that is 16.70 and 23.80 % higher than control and percent increase improvements of cotton shoot/root length with Br5 i.e. 18.3, 24.8 % higher than that of control. Higher values of IAA equivalents were observed in root/shoot content of cotton with isolate of Br5. Bacterial inoculation improved the plant height, boll weight, number of bolls plant-1 and seed cotton yield with Br5 inoculation. The chlorophyll content, photosynthetic rate, transpiration rate and photo active radiation were also higher in the inoculated treatments. Results of present study clearly demonstrated that different isolates of Rhizobium species improved the growth and yield parameters of cotton and thus Rhizobium sp can be effectively utilized as bacterial inoculants in non-legumes

The effects of long-term diabetes on the haematological and uterine indicators and their association with neonatal nephrogenesis counter-protected by camel milk: A time dependent study

The novelty of this project is to describe how chronic diabetes altered the haematological and uterine indicators in a time dependent-manner that were reversed by camel milk (CM) therapy in pregnant and nonpregnant rat models. Fifty-four female rats were divided into three groups: Placebo (N), diabetic control (DC) and diabetic treated (DT) with CM at 40 ml/kg/24 h for 90 days. A single intact male was introduced into every group for mating at day 60 of the experiment. The sample collection was undertaken at day 30 and 60 of the nonpregnant rats and at day 90 immediately after parturition for the pregnant rats. At every collection, the dam’s blood, as well as the uteri and neonatal kidneys were collected and subjected to a paraffin tissue preparation technique for a histological evaluation. The data revealed that at day 30, the uterine endo- and myometrium remained unaffected by diabetes, but at day 60, a significant reduction in the uterine indicators from diabetes was observed. However, the CM restored the uterine histology in the DT. At 90 day, chronic diabetes showed (P < 0.05) a harmful effect on the pregnant uterus which was reversed (P < 0.05) by the CM. The RBC (red blood cell) indices, platelets, and leucocyte counts were severely affected by the diabetes and protected by the CM at every point of collection. The kidney tissues of the neonate rats, delivered by the dams, in the DC presented a significant (P < 0.05) shrinkage in the cortex and glomeruli while the CM potentially reversed these changes. These results will help to understand the chronic diabetes effects on the uterus and neonate’s renal genesis, and the role of camel milk in the management of chronic pre-gestational diabete

The effects of long-term diabetes on the haematological and uterine indicators and their association with neonatal nephrogenesis counter-protected by camel milk: A time dependent study

The novelty of this project is to describe how chronic diabetes altered the haematological and uterine indicators in a time dependent-manner that were reversed by camel milk (CM) therapy in pregnant and nonpregnant rat models. Fifty-four female rats were divided into three groups: Placebo (N), diabetic control (DC) and diabetic treated (DT) with CM at 40 ml/kg/24 h for 90 days. A single intact male was introduced into every group for mating at day 60 of the experiment. The sample collection was undertaken at day 30 and 60 of the nonpregnant rats and at day 90 immediately after parturition for the pregnant rats. At every collection, the dam’s blood, as well as the uteri and neonatal kidneys were collected and subjected to a paraffin tissue preparation technique for a histological evaluation. The data revealed that at day 30, the uterine endo- and myometrium remained unaffected by diabetes, but at day 60, a significant reduction in the uterine indicators from diabetes was observed. However, the CM restored the uterine histology in the DT. At 90 day, chronic diabetes showed (P < 0.05) a harmful effect on the pregnant uterus which was reversed (P < 0.05) by the CM. The RBC (red blood cell) indices, platelets, and leucocyte counts were severely affected by the diabetes and protected by the CM at every point of collection. The kidney tissues of the neonate rats, delivered by the dams, in the DC presented a significant (P < 0.05) shrinkage in the cortex and glomeruli while the CM potentially reversed these changes. These results will help to understand the chronic diabetes effects on the uterus and neonate’s renal genesis, and the role of camel milk in the management of chronic pre-gestational diabete