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Background: Despite the short- and long-term acknowledged benefits of breastfeeding for mothers and their infants, worldwide rates trail behind international goals. Prior research confirms that breastfeeding is a nurse sensitive indicator and that problems with latching the baby and painful breastfeeding rank high among the reasons given for not continuing to breastfeed. The Lactation Assessment Tool (LATTM) was previously evaluated in a study conducted in Latvia by nurse midwives. Use of the LAT to assess breastfeeding and suggesting corrective interventions were shown to decrease pain and promote healing in traumatized nipples. The inter-rater reliability for that study was by test/re-test amongst participating researcher midwives. The aim of the current study is to expand the understanding of LAT inter-rater reliability to include novice and expert assessors. Methods: A convenience sample of twenty participants, including both novices (nine nursing students) and 11 self-identified experts, assessed four videos of breastfeeding dyads using the assessment tool, the LAT. Novice participants received a 2 h training session before final tool assessment. Each video was viewed three times, with a 3 min pause between viewings. All elements of the LAT that could be visually evaluated were included, with each element appearing in at least two of the videos. Results: Acceptable internal consistency of the LAT tool was found, with Cronbach’s alpha measuring 0.799, 0.740, 0.756 and 0.735 for each video, respectively. The reliability of the novice assessors improved over the course of the four videos, from 0.484 and 0.610 to 0.714 and 0.711. All of the experts had Cronbach’s alpha numbers that were acceptable, ranging from 0.769 to 1.00. Conclusions: Results indicate that experts perform much better using the tool than trained novices. However, the subsequent use of the tool resulted in the last two video assessments having an acceptable measure for the trained novice group. The LAT is a reliable tool for trained novices and experts to assess breastfeeding positioning and latch.

Cetylpyridinium chloride (CPC) is a quaternary ammonium antimicrobial used in numerous personal care products, human food, cosmetic products, and cleaning solutions. Yet, there is minimal published data on CPC effects on eukaryotes, immune signaling, and human health. Previously, it was shown that low-micromolar CPC inhibits rat mast cell function by inhibiting antigen (Ag)-stimulated Ca2+ mobilization, microtubule polymerization, and degranulation. In the current study, these findings are extended to human mast cells (LAD2); this paper presents data indicating that a mechanism of action for CPC might center on its positively-charged quaternary nitrogen in its pyridinium headgroup. The inhibitory effect of CPC was independent of signaling platform receptor architecture. Tyrosine phosphorylation events are a trigger of Ca2+ mobilization necessary for degranulation. CPC inhibits global tyrosine phosphorylation in Ag-stimulated mast cells. Specifically, CPC inhibits tyrosine phosphorylation of specific key players Syk kinase and LAT, a substrate of Syk. In contrast, CPC did not affect Lyn kinase phosphorylation. Thus, a root mechanism for CPC effect might be electrostatic disruption of particular tyrosine phosphorylation events essential for signaling. This work presented here outlines biochemical mechanisms underlying the effects of CPC on immune signaling.

Human alphaherpesvirus 1 (HSV-1) establishes life-long latency in sensory neurons in trigeminal ganglia (TG), brainstem neurons, and other CNS neurons. Two important segments of the brainstem were examined in this study: principal sensory nucleus of the spinal trigeminal tract (Pr5) because it receives direct afferent inputs from TG, and locus coeruleus (LC) because it is indirectly connected to Pr5 and LC sends axonal projections to cortical structures, which may facilitate viral spread from brainstem to the brain. The only viral gene abundantly expressed during latency is the latency associated transcript (LAT). Previous studies revealed 8-week old female C57Bl/6 mice infected with a LAT null mutant (dLAT2903) versus wild-type (wt) HSV-1 exhibit higher levels of senescence markers and inflammation in LC of females. New studies revealed 1-year old mice latently infected with wt HSV-1 or dLAT2903 contained differences in neuroinflammation and senescence in Pr5 and LC versus young mice. In summary, these studies confirm HSV-1 promotes neuro-inflammation in the brainstem, which may accelerate neurodegenerative disease.

Introduction: Patients with oligometastatic NSCLC benefit from locally ablative therapies (LAT); the role of adjuvant systemic therapies, however, remains less clear. In a single-arm, phase II clinical trial, we found that patients with oligometastatic NSCLC treated with a year of pembrolizumab after LAT had superior progression-free survival (PFS) compared with a historical control cohort. Herein, we present long-term follow-up on PFS and overall survival (OS). Methods: From February 1, 2015, to September 30, 2017, 45 patients with synchronous or metachronous oligometastatic (≤4 metastatic sites) NSCLC treated with LAT to all sites received adjuvant pembrolizumab every 21 days for up to 16 cycles. The primary efficacy end point was PFS from the start of pembrolizumab. Secondary end points included OS and safety. Median duration of follow-up was 66 months, and data cutoff was December 1, 2022. Results: A total of 45 patients were enrolled and treated with pembrolizumab after LAT (median age, 64 y [range, 46–82]; 21 women [47%]; 31 with a solitary oligometastatic site [69%]). At the data cutoff, 32 patients had progressive disease, 19 patients had died, and 13 patients had no evidence of relapse. Median PFS was 19.7 months (95% confidence interval: 7.6–31.7 mo); median OS was not reached (95% confidence interval: 37.7 mo–not reached). OS at 5 years was 60.0% (SE, 7.4%). Metachronous oligometastatic disease was associated with improved OS and PFS through Cox proportional hazard models. Conclusions: Pembrolizumab after LAT for oligometastatic NSCLC results in promising PFS and OS with a tolerable safety profile.

The determination of the maritime boundary is related to the selection of the tidal datum used in relation to the zero-depth reference. This research was conducted to determine the impact of the zero-depth reference that has been used, the Mean Low Water Spring (MLWS), against the zero-depth recommended by the International Hydrographic Organization (IHO), the Lowest Astronomical Tide (LAT). Tidal data is obtained from BIG's tidal station processing, while the MLWS value data is obtained from a combination of analysis from the Hydrographic Office of Indonesia (Pushidros). Based on these data, it is found that 25 out of 37 tidal stations indicate that the LAT value is below the MLWS in its vertical position. This shows that sea boundary claims can potentially be shifted by changing the zero-depth reference from the original by switching from MLWS to LAT. Lastly, this study presents the benefits of implementing the vertical system reference recommended by the IHO, the LAT, as an international chart datum.

Abstract Background Toxoplasma is an obligate intracellular protozoan that causes an important zoonotic disease with a worldwide distribution. Felids are the definitive hosts of this parasite, while virtually all warm-blooded animals, including birds, serve as intermediate hosts. Four ring-tailed lemurs (Lemur catta) in the Taipei Zoo died of acute Toxoplasma infection in June 2019. Since then, Toxoplasma has occasionally been identified in this Zoo during necropsy of dead animals and PCR of animal blood samples. Therefore, a general survey of Toxoplasma infection in animals in the Zoo seems to be needed. Methods and results An indirect multispecies ELISA was used for the first time to screen for Toxoplasma infection in 326 serum samples collected from 75 species of animals. The infection rate of Toxoplasma was 27% (88/326). A commercial latex agglutination (LAT) assay was used to re-examine the samples with doubtful and uncertain ELISA results (151 samples from 42 species). The infection rate increased to 36.2% (118/326), and the indirect multispecies ELISA appeared to be applicable to 31 of 75 species animals included in this study. Nested PCR assays targeting the dense granule protein 7 (GRA7) gene and B1 gene were also used to detect Toxoplasma in DNA samples extracted from 10 liver or blood specimens from 8 animals. GRA7 gene fragments were amplified from 8 samples from 7 animals, while B1 gene fragments were amplified from only 4 samples from 4 animals. From the B1 nested PCR and the sequence data of GRA7 fragments amplified from infectious specimens, the animals in the Zoo were speculated to have been infected by at least three different Toxoplasma variants. Conclusions According to the serological investigation, we speculated that over one-third (36.2%) of animals in Taipei Zoo presented the infection of Toxoplasma, and the indirect multispecies ELISA we used can be applied to detect Toxoplasma infection in 31 animal species included in this study. Sequence analysis revealed that at least three Toxoplasma variants were infecting the animals of Taipei Zoo.

Specific antigen recognition is one of the immune system’s features that allows it to mount intense yet controlled responses to an infinity of potential threats. T cells play a relevant role in the host defense and the clearance of pathogens by means of the specific recognition of peptide antigens presented by antigen-presenting cells (APCs), and, to do so, they are equipped with a clonally distributed antigen receptor called the T-cell receptor (TCR). Upon the specific engagement of the TCR, multiple intracellular signals are triggered, which lead to the activation, proliferation and differentiation of T lymphocytes into effector cells. In addition, this signaling cascade also operates during T-cell development, allowing for the generation of cells that can be helpful in the defense against threats, as well as preventing the generation of autoreactive cells. Early TCR signals include phosphorylation events in which the tyrosine kinases Lck and ZAP70 are involved. The sequential activation of these kinases leads to the phosphorylation of the transmembrane adaptor LAT, which constitutes a signaling hub for the generation of a signalosome, finally resulting in T-cell activation. These early signals play a relevant role in triggering the development, activation, proliferation and apoptosis of T cells, and the negative regulation of these signals is key to avoid aberrant processes that could generate inappropriate cellular responses and disease. In this review, we will examine and discuss the roles of the tyrosine kinases Lck and ZAP70 and the membrane adaptor LAT in these cellular processes.

Toxoplasmosis is an important worldwide foodborne zoonotic disease. Infected cattle meats is considered a serous cause of human toxoplasmosis. Here, this study assesses the infection with Toxoplasma gonddi (T. gondii) in cattle using meat juice samples from diaphragmatic muscles collected at the slaughter. An in house indirect enzyme linked immunosorbent assay (ELISA), and commercial latex agglutination test (LAT) followed by immunoblotting were developed on the meat juice (fluids) using tachyzoites of locally isolated T. gondii strain. The comparative analysis of the results of the tested juice samples showed an excellent agreement between the in-house indirect ELISA and LAT test in the positive and negative of meat juice. Relative sensitivity was higher for ELISA on diaphragms fluids random samples 80.39%, for the LAT test was 68.6%. Immune-reactive bands of T. gondii local strain Ag with naturally infected meat juice were 116, 83, 65, 30 and 23 KDa. The obtained results concluded that the development of an effective ELISA test to be used in for detection of toxoplasmosis infection of slaughtered cattle in large-scale would be exactly valuable, since the important role that beef plays in epidemiology of T. gondii, in particular the hazard of transmission to human and food safety.

Toxoplasma gondii is a protozoan parasite prevalent in humans and other animals worldwide having medical and veterinary importance on account of reproductive failure causing significant socioeconomic losses. The aims of this study were to estimate the seroprevalence of T. gondii infection in goats, determined the possible risk factors associated, and evaluate the performances of the latex agglutination test (LAT) to anti-T. gondii antibodies screening using the indirect Enzyme-linked immunosorbent assay as a reference test (iELISA). A total of 184 serum samples from goats reared on 25 farms in Mila district from North-Eastern Algeria were collected and tested for anti-T. gondii IgG antibodies using two commercial serological tests (ELISA and LAT). A seroprevalence rate of 71.73 % and 63.58 % was obtained by both ELISA and LAT tests, respectively. The analysis of some factors thought to be related to the onset of this infection such as age, sex, management system, locality and presence of cats showed no significant relationship (P > 0.05); these factors did not seem to affect the frequency of the infection. The seropositivity level of T. gondii was significantly higher in aborted goats (P = 0.007), which suggested that they may play a significant role in pregnancy failure. In the concordance evaluation between the two serological tests (ELISA and LAT), the Cohen’s Kappa value was calculated and the results showed a K of 0.519 (p = 0.000) belonging to the range of 0.41—0.60 indicating just average agreement. The results of the Mc Nemar test showed that both tests gave significantly different results and seropositivity values (P < 0.05). The high prevalence observed in this study indicated a widespread exposure to T. gondii from goats and the potential risk of T. gondii infection for humans in North-Eastern Algeria. These results elucidate the challenges of applying serology to estimate goat exposure to T. gondii. The choice between the two serological tests will depend on their performances, as well as the availability of the equipment, laboratory conditions and the number of samples to be tested.

The aim of this work is to verify the possibility of transport of 26 biologically active ultrashort peptides (USPs) into cells via LAT and PEPT family transporters. Molecular modeling and computer-assisted docking of peptide ligands revealed that the size and structure of ligand-binding sites of the amino acid transporters LAT1, LAT2, and of the peptide transporter PEPT1 are sufficient for the transport of the 26 biologically active di-, tri-, and tetra-peptides. Comparative analysis of the binding of all possible di- and tri-peptides (8400 compounds) at the binding sites of the LAT and PEPT family transporters has been carried out. The 26 biologically active USPs systematically showed higher binding scores to LAT1, LAT2, and PEPT1, as compared with di- and tri-peptides, for which no biological activity has been established. This indicates an important possible role which LAT and PEPT family transporters may play in a variety of biological activities of the 26 biologically active peptides under investigation in this study. Most of the 26 studied USPs were found to bind to the LAT1, LAT2, and PEPT1 transporters more efficiently than the known substrates or inhibitors of these transporters. Peptides ED, DS, DR, EDR, EDG, AEDR, AEDL, KEDP, and KEDG, and peptoids DS7 and KE17 with negatively charged Asp− or Glu− amino acid residues at the N-terminus and neutral or positively charged residues at the C-terminus of the peptide are found to be the most effective ligands of the transporters under investigation. It can be assumed that the antitumor effect of the KE, EW, EDG, and AEDG peptides could be associated with their ability to inhibit the LAT1, LAT2, and PEPT1 amino acid transporters. The data obtained lead to new prospects for further study of the mechanisms of transport of USP-based drugs into the cell and design of new antitumor drugs.

Livestock meat is considered a potential source of human infection, therefore evaluation of the infection rate with Toxoplasma gondii (T. gondii) in these animals’ meat is needed for public health protection. Serum and meat samples were collected from slaughtered animals (n=106) (27 sheep, 42 cattle and 37 camels) from slaughterhouses in Aswan Governorate, located in southern Egypt. T. gondii infection status was detected by Latex agglutination test (LAT), microscopic examination and PCR. Our results revealed that the overall prevalence of anti-T. gondii antibodies among serum samples was 48.1 % using LAT. Male sheep, cattle, and camels had a higher positive percentage (42.9 %, 69.4% and 33.3 %, respectively) than females (40 %, 50 % and 30.8 %, respectively). The infection rate of T. gondii was higher in adult sheep and camels, while it was lower in adult cattle (64.5 %) than in young (72.7 %). There was a high statistically significant difference (P ≤ 0.01) between the animal species using LAT. T. gondii trophozoites were detected microscopically in 47.1 % of meat samples, while T. gondii DNA was detected in 56.9 % by using conventional Polymerase Chain Reaction (cPCR) based B1 gene. There was no significant difference between the results of both microscopy and PCR methods. In conclusion, PCR outperforms LAT in the detection of T. gondii infection and can thus be applied to routine diagnosis of toxoplasmosis in red meat producing animals.

Adaptor molecules lack enzymatic and transcriptional activities. Instead, they exert their function by linking multiple proteins into intricate complexes, allowing for transmitting and fine-tuning of signals. Many adaptor molecules play a crucial role in T-cell signaling, following engagement of the T-cell receptor (TCR). In this review, we focus on Linker of Activation of T cells (LAT) and SH2 domain-containing leukocyte protein of 76 KDa (SLP-76). Monogenic defects in these adaptor proteins, with known roles in T-cell signaling, have been described as the cause of human inborn errors of immunity (IEI). We describe the current knowledge based on defects in cell lines, murine models and human patients. Germline mutations in Adhesion and degranulation adaptor protein (ADAP), have not resulted in a T-cell defect.

Self- and non-self ligand discrimination is a core principle underlying T cell-mediated immunity. Mature αβ T cells can respond to a foreign peptide ligand presented by major histocompatibility complex molecules (pMHCs) on antigen presenting cells, on a background of continuously sensed self–pMHCs. How αβ T cells can properly balance high sensitivity and high specificity to foreign pMHCs, while surrounded by a sea of self-peptide ligands is not well understood. Such discrimination cannot be explained solely by the affinity parameters of T cell antigen receptor (TCR) and pMHC interaction. In this review, we will discuss how T cell ligand discrimination may be molecularly defined by events downstream of the TCR–pMHC interaction. We will discuss new evidence in support of the kinetic proofreading model of TCR ligand discrimination, and in particular how the kinetics of specific phosphorylation sites within the adaptor protein linker for activation of T cells (LAT) determine the outcome of TCR signaling. In addition, we will discuss emerging data regarding how some kinases, including ZAP-70 and LCK, may possess scaffolding functions to more efficiently direct their kinase activities.

Toxoplasma gondii is a protozoan parasite that causes toxoplasmosis in warm-blooded vertebrates, globally. The main aims of this study were to assess the seropositivity to toxoplasmosis of an exotic breed of cattle (n = 400) from different farms using the Latex Agglutination Test and validate Cattle Toxo IgG ELISA kit. Of a total of 400 cattle sera that were evaluated by LAT, 143 (35.75%) were found positive. Based on these data, 90 samples (n = 60 seronegative by LAT; n = 30 seropositive by LAT) were elected for screening through a commercially available ELISA kit. The same 90 samples were screened through a Cattle Toxo IgG ELISA kit for validation purposes. Of 90 samples, 40 were seropositive in the Cattle Toxo IgG ELISA kit (100% sensitivity), and 38 were seropositive in a commercially available ELISA kit. All 50 samples in the Cattle Toxo IgG ELISA kit (96.15% specificity) were also seronegative in the commercially available ELISA kit. Hence, the sensitivity and specificity of the Cattle Toxo IgG ELISA kit came out to be 100% and 96.15%, and in LAT, it was found as 26.31% and 61.53%, respectively. Therefore, the Cattle Toxo IgG ELISA kit is a highly reliable serodiagnostic tool to diagnose bovine toxoplasmosis.

The adaptor LAT plays a crucial role in the transduction of signals coming from the TCR/CD3 complex. Phosphorylation of some of its tyrosines generates recruitment sites for other cytosolic signaling molecules. Tyrosine 132 in human LAT is essential for PLC-γ activation and calcium influx generation. It has been recently reported that a conserved glycine residue preceding tyrosine 132 decreases its phosphorylation kinetics, which constitutes a mechanism for ligand discrimination. Here we confirm that a LAT mutant in which glycine 131 has been substituted by an aspartate (LATG131D) increases phosphorylation of Tyr132, PLC-γ activation and calcium influx generation. Interestingly, the LATG131D mutant has a slower protein turnover while being equally sensitive to Fas-mediated protein cleavage by caspases. Moreover, J.CaM2 cells expressing LATG131D secrete greater amounts of interleukin-2 (IL-2) in response to CD3/CD28 engagement. However, despite this increased IL-2 secretion, J.CaM2 cells expressing the LATG131D mutant are more sensitive to inhibition of IL-2 production by pre-treatment with anti-CD3, which points to a possible role of this residue in the generation of anergy. Our results suggest that the increased kinetics of LAT Tyr132 phosphorylation could contribute to the establishment of T cell anergy, and thus constitutes an earliest known intracellular event responsible for the induction of peripheral tolerance.

Global navigation satellite systems (GNSS) is widely utilized in marine applications for high accuracy vertical positioning. The errors introduced to charts by the present use of traditional shore-based discrete point chart datum (CD) can become a significant part of the total vertical error and deteriorates the vertical positioning accuracy estimated by GNSS technique. Therefore, a need for developing a CD in a continuous form is essential to replace the traditional CD so that it can be transformed from/to another continuous datum such as the reference ellipsoid (WGS84). The importance of the development of CD-to-WGS84 separation model is that it can be utilized by the hydrographers to develop an accurate vertical control for hydrographic surveys applications and can be utilized by the mariners to produce an accurate dynamic electronic navigation charts (ENC). In this paper, a Saudi continuous chart datum in the Arabian Gulf area is developed to provide CD-to-WGS84 separation model. It is shown that the Saudi continuous chart datum ranges from –14.96 m to –34.64 m and can be achieved with uncertainty ranges from 0.11 m to 0.16 m. The developed CD-to-WGS84 separation model in this paper can meet many maritime applications.

The human heart is an organ of high complexity and hence, very challenging to simulate. To calculate the force developed by the human heart and therefore the tension of the muscle fibers, accurate models are necessary. The force generated by the cardiac muscle has physiologically imposed limits and depends on various characteristics such as the length, strain and the contraction velocity of the cardiomyocytes. Another characteristic is the activation time of each cardiomyocyte, which is a wave and not a static value for all cardiomyocytes. To simulate a physiologically correct excitation, the functionality of the cardiac simulation framework CardioMechanics was extended to incorporate inhomogeneous activation times. The functionality was then used to evaluate the effects of local activation times with two different tension models. The active stress generated by the cardiomyocytes was calculated by (i) an explicit function and (ii) an ode-based model. The results of the simulations showed that the maximum pressure in the left ventricle dropped by 2.3% for the DoubleHill model and by 5.3% for the Lumens model. In the right ventricle the simulations showed similar results. The maximum pressure in both the left and the right atrium increased using both models. Given that the simulation of the inhomogeneously activated cardiomyocytes increases the simulation time when used with the more precise Lumens model, the small drop in maximum pressure seems to be negligible in favor of a simpler simulation model

The human heart is an organ of high complexity and hence, very challenging to simulate. To calculate the force developed by the human heart and therefore the tension of the muscle fibers, accurate models are necessary. The force generated by the cardiac muscle has physiologically imposed limits and depends on various characteristics such as the length, strain and the contraction velocity of the cardiomyocytes. Another characteristic is the activation time of each cardiomyocyte, which is a wave and not a static value for all cardiomyocytes. To simulate a physiologically correct excitation, the functionality of the cardiac simulation framework CardioMechanics was extended to incorporate inhomogeneous activation times. The functionality was then used to evaluate the effects of local activation times with two different tension models. The active stress generated by the cardiomyocytes was calculated by (i) an explicit function and (ii) an ode-based model. The results of the simulations showed that the maximum pressure in the left ventricle dropped by 2.3% for the DoubleHill model and by 5.3% for the Lumens model. In the right ventricle the simulations showed similar results. The maximum pressure in both the left and the right atrium increased using both models. Given that the simulation of the inhomogeneously activated cardiomyocytes increases the simulation time when used with the more precise Lumens model, the small drop in maximum pressure seems to be negligible in favor of a simpler simulation model.

A traditional shore-based discrete point chart datum (CD) that represents the lowest astronomical tide (LAT) in Saudi Arabia using tide gauge data is utilized to reduce the observed depth collected from hydrographic surveying test to CD-referenced depth for producing navigation charts for maritime navigation applications. A need for developing CD in a continuous form is essential to replace the traditional discrete CD using tide gauge data. The importance of the development of CD-to-ellipsoid (WGS84) separation model is that it can be utilized by the hydrographers to develop an accurate vertical control for hydrographic surveys applications and can be utilized by the mariners to produce accurate dynamic electronic navigation charts (ENCs). In this paper, a continuous CD to WGS84 ellipsoid separation model for the Sharm Obhur area is developed using a multibeam hydrographic surveying test. It is shown that the continuous chart datum ranges from −4.920 m to −4.766 m and can be achieved with standard deviation ranges from 0.1 cm to 2.3 cm. To validate the separation model, a comparison was made with the gravimetric/oceanographic method based on the separation height developed from geoid height, the sea surface topography and LAT value (chart datum to mean sea level) at the tide gauge located in the study area. The comparison showed that the average value of the developed continuous CD to WGS84 separation model heights using multibeam hydrographic surveying agrees with the separation height estimated from gravimetric/oceanographic method.

CD4+ T cells differentiate into T helper cell subsets in feedforward manners with synergistic signals from the T cell receptor (TCR), cytokines, and lineage-specific transcription factors. Naive CD4+ T cells avoid spontaneous engagement of feedforward mechanisms but retain a prepared state. T cells lacking the adaptor molecule LAT demonstrate impaired TCR-induced signals yet cause a spontaneous lymphoproliferative T helper 2 (TH2) cell syndrome in mice. Thus, LAT constitutes an unexplained maintenance cue. Here, we demonstrate that tonic signals through LAT constitutively export the repressor HDAC7 from the nucleus of CD4+ T cells. Without such tonic signals, HDAC7 target genes Nur77 and Irf4 are repressed. We reveal that Nur77 suppresses CD4+ T cell proliferation and uncover a suppressive role for Irf4 in TH2 polarization; halving Irf4 gene-dosage leads to increases in GATA3+ and IL-4+ cells. Our studies reveal that naive CD4+ T cells are dynamically tuned by tonic LAT-HDAC7 signals.

The modified agglutination test (MAT) has been widely used for the detection of Toxoplasma gondii infection in numerous animal species. For the standard protocol of MAT test, T. gondii whole cell antigens were produced in the laboratory mice, which was a tedious process. The produced antigen in cell culture was used to assure its capability for MAT test. For detecting the antibodies of T. gondii in human sera, comparison was made between MAT, enzyme linked immunosorbent assay (ELISA) and latex agglutination test (LAT). A total of 96 human serum samples were tested. The anti-Toxoplasma IgG antibodies were found in 25.0% (24/96), 20.8% (20/96) and 13.5% (13/96) samples by MAT (cutoff 1:25), ELISA and LAT tests, respectively. The MAT and ELISA tests matched 95.8% in detection of IgG antibodies, with the positive percent agreement of 100% (20/20) and negative percent agreement of 94.7% (72/76). For the MAT versus LAT tests, the overall agreement was 88.5% (85/96), with the positive percent agreement of 100% (13/13) and negative percent agreement of 86.7% (72/83). These results suggest a strong correlation between the MAT and ELISA tests in detecting serum IgG to T. gondii in human sera. In conclusion, T. gondii prepared in cell culture provides an alternative solution to produce antigens for MAT test.

Antigen receptor signaling pathways are organized by adaptor proteins. Three adaptors, LAT, Gads, and SLP-76, form a heterotrimeric complex that mediates signaling by the T cell antigen receptor (TCR) and by the mast cell high affinity receptor for IgE (FcεRI). In both pathways, antigen recognition triggers tyrosine phosphorylation of LAT and SLP-76. The recruitment of SLP-76 to phospho-LAT is bridged by Gads, a Grb2 family adaptor composed of two SH3 domains flanking a central SH2 domain and an unstructured linker region. The LAT-Gads-SLP-76 complex is further incorporated into larger microclusters that mediate antigen receptor signaling. Gads is positively regulated by dimerization, which promotes its cooperative binding to LAT. Negative regulation occurs via phosphorylation or caspase-mediated cleavage of the linker region of Gads. FcεRI-mediated mast cell activation is profoundly impaired in LAT- Gads- or SLP-76-deficient mice. Unexpectedly, the thymic developmental phenotype of Gads-deficient mice is much milder than the phenotype of LAT- or SLP-76-deficient mice. This distinction suggests that Gads is not absolutely required for TCR signaling, but may modulate its sensitivity, or regulate a particular branch of the TCR signaling pathway; indeed, the phenotypic similarity of Gads- and Itk-deficient mice suggests a functional connection between Gads and Itk. Additional Gads binding partners include costimulatory proteins such as CD28 and CD6, adaptors such as Shc, ubiquitin regulatory proteins such as USP8 and AMSH, and kinases such as HPK1 and BCR-ABL, but the functional implications of these interactions are not yet fully understood. No interacting proteins or function have been ascribed to the evolutionarily conserved N-terminal SH3 of Gads. Here we explore the biochemical and functional properties of Gads, and its role in regulating allergy, T cell development and T-cell mediated immunity.

In this paper we study the problem of recovering a secret S-box from its difference distribution table (DDT). While being an interesting theoretical problem on its own, the ability to recover the S-box from the DDT of a secret S-box can be used in cryptanalytic attacks where the attacker can obtain the DDT (e.g., in Bar-On et al.’s attack on GOST), in supporting theoretical analysis of the properties of difference distribution tables (e.g., in Boura et al.’s work), or in some analysis of S-boxes with unknown design criteria (e.g., in Biryukov and Perrin’s analysis). We show that using the well established relation between the DDT and the linear approximation table (LAT), one can devise an algorithm different from the straightforward guess-and-determine (GD) algorithm proposed by Boura et al. Moreover, we show how to exploit this relation, and embed the knowledge obtained from it in the GD algorithm. We tested our new algorithm on random S-boxes of different sizes, and for random 14-bit bijective S-boxes, our results outperform the GD attack by several orders of magnitude.

Intracellular signaling through the T cell receptor (TCR) is essential for T cell development and function. Proper TCR signaling requires the sequential activities of Lck and ZAP-70 kinases, which result in the phosphorylation of tyrosine residues located in the CD3 ITAMs and the LAT adaptor, respectively. LAT, linker for the activation of T cells, is a transmembrane adaptor protein that acts as a scaffold coupling the early signals coming from the TCR with downstream signaling pathways leading to cellular responses. The leukemic T cell line Jurkat and its derivative mutants J.CaM1.6 (Lck deficient) and J.CaM2 (LAT deficient) have been widely used to study the first signaling events upon TCR triggering. In this work, we describe the loss of LAT adaptor expression found in a subline of J.CaM1.6 cells and analyze cis-elements responsible for the LAT expression defect. This new cell subline, which we have called J.CaM1.7, can re-express LAT adaptor after Protein Kinase C (PKC) activation, which suggests that activation-induced LAT expression is not affected in this new cell subline. Contrary to J.CaM1.6 cells, re-expression of Lck in J.CaM1.7 cells was not sufficient to recover TCR-associated signals, and both LAT and Lck had to be introduced to recover activatory intracellular signals triggered after CD3 crosslinking. Overall, our work shows that the new LAT negative J.CaM1.7 cell subline could represent a new model to study the functions of the tyrosine kinase Lck and the LAT adaptor in TCR signaling, and their mutual interaction, which seems to constitute an essential early signaling event associated with the TCR/CD3 complex.

LAT is an important player of the signaling cascade induced by TCR activation. This adapter molecule is present at the plasma membrane of T lymphocytes and more abundantly in intracellular compartments. Upon T cell activation the intracellular pool of LAT is recruited to the immune synapse (IS). We previously described two pathways controlling LAT trafficking: retrograde transport from endosomes to the TGN, and anterograde traffic from the Golgi to the IS. We address the specific role of four proteins, the GTPase Rab6, the t-SNARE syntaxin-16, the v-SNARE VAMP7 and the golgin GMAP210, in each pathway. Using different methods (endocytosis and Golgi trap assays, confocal and TIRF microscopy, TCR-signalosome pull down) we show that syntaxin-16 is regulating the retrograde transport of LAT whereas VAMP7 is regulating the anterograde transport. Moreover, GMAP210 and Rab6, known to contribute to both pathways, are in our cellular context, specifically and respectively, involved in anterograde and retrograde transport of LAT. Altogether, our data describe how retrograde and anterograde pathways coordinate LAT enrichment at the IS and point to the Golgi as a central hub for the polarized recruitment of LAT to the IS. The role that this finely-tuned transport of signaling molecules plays in T-cell activation is discussed.

Among various types of membrane proteins that are regulated by cytoskeleton, the T cell receptor (TCR) greatly benefits from these cellular machineries for its function. The T cell is activated by the ligation of TCR to its target agonist peptide. However, the binding affinity of the two is not very strong, while the T cell needs to discriminate agonist from many nonagonist peptides. Moreover, the strength and duration of the activation signaling need to be tuned for immunological functions. Many years of investigations revealed that dynamic acto-myosin cytoskeletons and plasma membranes in T cells facilitate such regulations by modulating the spatiotemporal distributions of proteins in plasma membranes and by applying mechanical loads on proteins. In these processes, protein dynamics in multiple scales are involved, ranging from collective molecular motions and macroscopic molecular organizations at the cell–cell interface to microscopic changes in distances between receptor and ligand molecules. In this review, details of how cytoskeletons and membranes regulate these processes are discussed, with the emphasis on how all these processes are coordinated to occur within a single cell system.

The Arabidopsis L-type Amino Acid Transporter-5 (LAT5; At3g19553) was recently studied for its role in developmental responses such as flowering and senescence, under an assumption that it is a polyamine uptake transporter (PUT5). The LATs in Arabidopsis have a wide range of substrates, including amino acids and polyamines. This report extensively studied the organ and tissue-specific expression of the LAT5/PUT5 and investigated its role in mediating amino acid transport. Organ-specific quantitative RT-PCR detected LAT5/PUT5 transcripts in all organs with a relatively higher abundance in the leaves. Tissue-specific expression analysis identified GUS activity in the phloem under the LAT5/PUT5 promoter. In silico analysis identified both amino acid transporter and antiporter domains conserved in the LAT5/PUT5 protein. The physiological role of the LAT5/PUT5 was studied through analyzing a mutant line, lat5-1, under various growth conditions. The mutant lat5-1 seedlings showed increased sensitivity to exogenous leucine in Murashige and Skoog growth medium. In soil, the lat5-1 showed reduced leaf growth and altered nitrogen content in the seeds. In planta radio-labelled leucine uptake studies showed increased accumulation of leucine in the lat5-1 plants compared to the wild type when treated in the dark prior to the isotopic feeding. These studies suggest that LAT5/PUT5 plays a role in mediating amino acid transport.

Developing an accurate Lowest Astronomical Tide (LAT) in a continuous form is essential for many maritime applications as it can be employed to develop an accurate continuous vertical control datum for hydrographic surveys applications and to produce accurate dynamic electronic navigation charts for safe maritime navigation by mariners. The LAT can be developed in a continuous (surface) using an estimated LAT surface model from the hydrodynamic ocean model along with coastal discrete LAT point values derived from tide gauges data sets to provide the corrected LAT surface model. In this paper, an accurate LAT surface model was developed for the Red Sea case study using a Maximum Likelihood Estimator (MLE) with multiple hydrodynamic ocean models hybridization, namely, WebTide, FES2014, DTU10, and EOT11a models. It was found that the developed optimal hybrid LAT model using MLE with multiple hydrodynamic ocean models hybridization ranges from 0.1 m to 1.63 m, associated with about 2.4 cm of uncertainty at a 95% confidence level in the Red Sea case study area. To validate the accuracy of the developed model, the comparison was made between the optimal hybrid LAT model developed from multiple hydrodynamic ocean models hybridization using the MLE method with the individual LAT models estimated from individual WebTide, FES2014, DTU10, or EOT11a ocean models based on the associated uncertainties estimated at a 95% confidence level. It was found that the optimal hybrid LAT model accuracy is superior to the individual LAT models estimated from individual ocean models with an improvement of about 50% in average, based on the estimated uncertainties. The importance of developing optimal LAT surface model using the MLE method with multiple hydrodynamic ocean models hybridization in this paper with few centimeters level of uncertainty can lead to accurate continuous vertical datum estimation that is essential for many maritime applications.

The adaptor protein linker for activation of T cells (LAT) has an essential role transducing activatory intracellular signals coming from the TCR/CD3 complex. Previous reports have shown that upon T-cell activation, LAT interacts with the tyrosine kinase Lck, leading to the inhibition of its kinase activity. LAT–Lck interaction seemed to depend on a stretch of negatively charged amino acids in LAT. Here, we have substituted this segment of LAT between amino acids 113 and 126 with a non-charged segment and expressed the mutant LAT (LAT-NIL) in J.CaM2 cells in order to analyze TCR signaling. Substitution of this segment in LAT prevented the activation-induced interaction with Lck. Moreover, cells expressing this mutant form of LAT showed a statistically significant increase of proximal intracellular signals such as phosphorylation of LAT in tyrosine residues 171 and 191, and also enhanced ZAP70 phosphorylation approaching borderline statistical significance (p = 0.051). Nevertheless, downstream signals such as Ca2+ influx or MAPK pathways were partially inhibited. Overall, our data reveal that LAT–Lck interaction constitutes a key element regulating proximal intracellular signals coming from the TCR/CD3 complex.

Herpes simplex virus 1 (HSV1) is an important pathogen with a worldwide epidemic trend that affects populations of various ages. It has a high morbidity, particularly in juveniles, but a successful HSV1 vaccine is not currently available. Thus, our study systematically observed the immune responses induced in mice immunized with the attenuated HSV1 M3 mutant strain, which has mutations in the genes encoding the UL7 and Vhs tegument proteins and the latency-associated transcript. The immunity induced by the M3 mutant strain can control acute viral infection during HSV1 wild-type strain infection. Moreover, this immunity exerts a potent effect on controlling viral entry into the trigeminal neurons. These data encourage further studies investigating the development of M3 as a potential vaccine candidate, and much work is necessary to evaluate the safety and improve the immunogenicity of this strain.

This study was conducted to investigate the anti-toxoplasma antibodies titer (IgG and IgM), in aborted ewes in different localities of Duhok Governorate. The study was carried out in the laboratory of post graduated study, Faculty of Veterinary Medicine, Duhok University from October 2010 to April 2011. Four hundred and ninety-six (496) serum samples of aborted ewes (98, 144, 152 and 102 were obtained from Aqra, Duhok center, Shikhan and Zakho, respectively). Latex Agglutination test (LAT), Modified agglutination test (MAT) and Enzyme Linked Immunosorbent Assay (ELISA) were used for the aforementioned purpose. The over all prevalence of toxoplasmosis was (83.3%), and the rate was different in different localities such as Aqra (86.7%), Duhok (89.6%), Shikhan (74.3%) and Zakho (84.3%) using LAT. Out of 413 sera, sample which were confirmed positively LAT (67.31%) were positive for Toxoplasma by MAT (have had IgG type of antibody) and 149 (36.08%) sera gave positive reaction by IgM ELISA test (have had IgM type of antibody). The result revealed that the rate in different age groups was statistically different and the rate was 90.38% in age group

The sequence of events that initiates T cell signaling is dictated by the specificities and order of activation of the tyrosine kinases that signal downstream of the T cell receptor. Using a platform that combines exhaustive point-mutagenesis of peptide substrates, bacterial surface-display, cell sorting, and deep sequencing, we have defined the specificities of the first two kinases in this pathway, Lck and ZAP-70, for the T cell receptor ζ chain and the scaffold proteins LAT and SLP-76. We find that ZAP-70 selects its substrates by utilizing an electrostatic mechanism that excludes substrates with positively-charged residues and favors LAT and SLP-76 phosphosites that are surrounded by negatively-charged residues. This mechanism prevents ZAP-70 from phosphorylating its own activation loop, thereby enforcing its strict dependence on Lck for activation. The sequence features in ZAP-70, LAT, and SLP-76 that underlie electrostatic selectivity likely contribute to the specific response of T cells to foreign antigens.

AbstractThere is emerging evidence that exocytosis plays an important role in regulating T cell receptor (TCR) signaling. The trafficking molecules involved in lytic granule (LG) secretion in cytotoxic T lymphocytes (CTL) have been well studied due to the immune disorder known as familial hemophagocytic lymphohisiocytosis (FHLH). However, the knowledge of trafficking machineries regulating the exocytosis of receptors and signaling molecules remains quite limited. In this review, we summarize the reported trafficking molecules involved in the transport of the TCR and downstream signaling molecules to the cell surface. By combining this information with the known knowledge of LG exocytosis and general exocytic trafficking machinery, we attempt to draw a more complete picture of how the TCR signaling network and exocytic trafficking matrix are interconnected to facilitate T cell activation. This also highlights how membrane compartmentalization facilitates the spatiotemporal organization of cellular responses that are essential for immune functions.

The plasma membrane contains diverse and specialized membrane domains, which include tetraspanin-enriched domains (TEMs) and transmembrane adaptor protein (TRAP)-enriched domains. Recent biophysical, microscopic and functional studies indicated that TEMs and TRAP-enriched domains are involved in compartmentalization of physicochemical events of such important processes as immunoreceptor signal transduction and chemotaxis. Moreover, there is evidence of a cross-talk between TEMs and TRAP-enriched domains. In this review we discuss the presence and function of such domains and their crosstalk using mast cells as a model. The combined data based on analysis of selected mast cell-expressed tetraspanins [cluster of differentiation (CD)9, CD53, CD63, CD81, CD151)] or TRAPs [linker for activation of T cells (LAT), non-T cell activation linker (NTAL), and phosphoprotein associated with glycosphingolipid-enriched membrane microdomains (PAG)] using knockout mice or specific antibodies point to a diversity within these two families and bring evidence of the important roles of these molecules in signaling events. An example of this diversity is physical separation of two TRAPs, LAT and NTAL, which are in many aspects similar but show plasma membrane location in different microdomains in both non-activated and activated cells. Although our understanding of TEMs and TRAP-enriched domains is far from complete, pharmaceutical applications of the knowledge about these domains are under way.

Introduction: The cases of cryptococcal meningitis and other forms of cryptococcosis have increased in recent time and the present scenario of the condition with significant morbidity and mortality is actually posing a serious threat to the community, so an early and prompt diagnosis is necessary to prevent serious complications and thus improving the overall disease outcome. Aim: Comparison of diagnostic efficacy of nested Polymerase Chain Reaction (PCR) with Latex Agglutination Test (LAT) in the Cerebro Spinal Fluid (CSF) samples of the cases of meningitis in HIV positive and negative cases. Materials and Methods: We have compared the diagnostic efficacy of Latex Agglutination Test (LAT) with nested Polymerase Chain Reaction (PCR) in 200 Cerebrospinal Fluid (CSF) samples, including 14 HIV positive also, in the cases of suspected cryptococcal meningitis. Nested PCR was done in all cases reporting positive by LAT and results were then compared with that of India ink and culture on Sabouraud Dextrose Agar (SDA), and the isolates were further identified by urease, nitrate and sugar assimilation tests. Results: Of the 200 cases, including 14 HIV positive, LAT was positive in 46 cases while 154 were negative. Out of these 46 LAT positive cases, nested PCR was positive in 40 cases only, while culture and India ink was positive in 38 and 33 cases respectively. Majority of the cases, 30 (65.2%) were between age group 21-50 years, while 2 (4.3%) in 0-20, and 14 (30.4%) in 51-80 years age group. Conclusion: Although negative staining like India ink and nigrosin are most widely used techniques, but these suffer with subjective error. Rapid method like LAT is available but it always has the scope of false positive and negative results. In such cases nested PCR can help in establishing final diagnosis.

SIMON and SPECK families of block ciphers are well-known lightweight ciphers designed by the NSA. In this note, based on the previous investigations on SIMON, a closed formula for the squared correlations and differential probabilities of the mapping ϕ ( x ) = x ⊙ S 1 ( x ) on F 2 n is given. From the aspects of linear and differential cryptanalysis, this mapping is equivalent to the core quadratic mapping of SIMON via rearrangement of coordinates and EA -equivalence. Based on the proposed explicit formula, a full description of DDT and LAT of ϕ is provided. In the case of SPECK, as the only nonlinear operation in this family of ciphers is addition mod 2 n , after reformulating the formula for linear and differential probabilities of addition mod 2 n , straightforward algorithms for finding the output masks with maximum squared correlation, given the input masks, as well as the output differences with maximum differential probability, given the input differences, are presented. By the aid of the tools given in this paper, the process of the search for linear and differential characteristics of SIMON and SPECK families of block ciphers could be sped up, and the complexity of linear and differential attacks against these ciphers could be reduced.

Herpes simplex virus type-1 (HSV-1) initially infects mucoepithelial tissues of the orofacial region, the eye and to a lesser extent the genitalia. Subsequently, the virus is retrogradely transported through the axons of the sensory and sympathetic neurons to their nuclei, where the virus establishes a life-long latent infection. During this latency period, the viral genome is transcriptionally silent except for a single region encoding the latency-associated transcript (LAT). LAT has been shown to affect apoptosis, but little else is known regarding its effects on neurons. To understand how HSV-1 latency might affect dendrites in sympathetic neurons, we transfected primary cultures of sympathetic neurons obtained from rat embryos, with LAT expressing plasmids. LAT inhibited initial dendritic growth and induced dendritic retraction in sympathetic neurons. Latent HSV-1 infection of cultured sympathetic neurons inhibited dendritic growth indicating that this is likely also a consequence of natural infection.

The Ras-MAPK signaling pathway is highly conserved throughout evolution and is activated downstream of a wide range of receptor stimuli. Ras guanine nucleotide exchange factors (RasGEFs) catalyze GTP loading of Ras and play a pivotal role in regulating receptor-ligand induced Ras activity. In T cells, three families of functionally important RasGEFs are expressed: RasGRF, RasGRP, and SOS-family GEFs.Early on it was recognized that Ras activation is critical for T cell development and that the RasGEFs play an important role herein. More recent work has revealed that nuances in Ras activation appear to significantly impact T cell development and selection. These nuances include distinct biochemical patterns of analog versus digital Ras activation, differences in cellular localization of Ras activation, and intricate interplays between the RasGEFs during distinct T cell developmental stages as revealed by various new mouse models. In many instances, the exact nature of these nuances in Ras activation or how these may result from fine-tuning of the RasGEFs is not understood.One large group of biomolecules critically involved in the control of Ras-GEFs´functions are lipid second messengers. Multiple, yet distinct lipid products are generated following T cell receptor (TCR) stimulation and bind to different domains in the RasGRP and SOS RasGEFs to facilitate the activation of the membrane-anchored Ras GTPases. In this review we highlight how different lipid-based elements are generated by various enzymes downstream of the TCR and other receptors and how these dynamic and interrelated lipid products may fine-tune Ras activation by RasGEFs in developing T cells.

Back ground: Toxoplasmosis is an important zoonosis that causes economic losses in animal herds due to abortion and stillbirth as well as changes in the reproductive and neural system of susceptible animals . Objective: The aims of the present study is to determination the prevalence of T. gondii in farm animals ( sheep& goat)of both genders and different ages in Sulaimani province by using two serological tests (ELISA and LAT). Methods: Blood samples were collected from farm animals ,142 sheep and 46 goats , of different sexes and ages. Tow different serological tests ,ELISA and LAT for qualitative determination of T. gondii antibody titer in sheep and goats serum samples. Results: The prevalence rate in sheep was 73 (51.7 %) and 82 (57 %) , and 21 (54.6 %) and 25 (54.35 %) in goats ,by ELISA and LAT respectively. The prevalence of toxoplasmosis was highest in age group 7-9 (66.6%) in sheep in compares’ with other age groups. There was no significant differences between both spp.and tow test. Conclusion: Statistical results show no significant differences between both tests (ELISA &LAT) at (P ≥ 0.05).The prevalence of toxoplasmosis was increased proportionally with the age of animals, while gender has no effect on the prevalent rate .

Background: Most research asks whether or not cohabitation has come to rival marriage. Little is known about the meaning of living apart together (LAT) relationships, and whether LAT is an alternative to marriage and cohabitation or a dating relationship. Objective: We examine across Europe: (1) the prevalence of LAT, (2) the reasons for LAT, and (3) the correlates of (a) LAT relationships vis-à-vis being single, married, or cohabiting, and (b) different types of LAT union. Methods: Using Generations and Gender Survey data from ten Western and Eastern European countries, we present descriptive statistics about LATs and estimate multinominal logistic regression models to assess the correlates of being in different types of LAT unions. Results: LAT relationships are uncommon, but they are more common in Western than Eastern Europe. Most people in LAT unions intend to live together but are apart for practical reasons. LAT is more common among young people, those enrolled in higher education, people with liberal attitudes, highly educated people, and those who have previously cohabited or been married. Older people and divorced or widowed persons are more likely to choose LAT to maintain independence. Surprisingly, attitudinal and educational differences are more pronounced in Eastern Europe than in Western Europe. Conclusions: A tentative conclusion is that LAT is more often a stage in the union formation process than an alternative to marriage and cohabitation. Yet some groups do view LAT as substituting for marriage and cohabitation, and these groups differ between East and West. In Eastern Europe a cultural, highly educated elite seems to be the first to resist traditional marriage norms and embrace LAT (and cohabitation) as alternative living arrangements, whereas this is less the case in Western Europe. In Western Europe, LAT unions are mainly an alternative for persons who have been married before or had children in a prior relationship.

We use two surveys to describe the demographic and attitudinal correlates of being in "Living Apart Together" (LAT), cohabiting, and marital relationships for heterosexuals, lesbians, and gay men. About one third of U.S. adults not married or cohabiting are in LAT relationships - these individuals would be classified as "single" in conventional studies that focus on residential unions. Gay men are somewhat more likely than heterosexual men to be in LAT relationships. For heterosexuals and lesbians, LAT relationships are more common among younger people. Heterosexuals in LAT unions are less likely to expect to marry their partners, but more likely to say that couples should be emotionally dependent than are cohabiters. Regardless of sexual orientation, people in LAT relationships perceive similar amounts of emotional support from partners, but less instrumental support than cohabiters perceive.

In Spain, nearly two-thirds of women aged 20-34 have not yet entered their first union. However, almost half of them have a stable partner in a different household. Hence, the drop in marriage rates and low prevalence of cohabitation cannot be rightly interpreted as a decline in partnership formation, but rather as a postponement of co-residential unions. This paper examines the prevalence and determinants of non-residential stable partnerships among young adults (women aged 20-34), in relation to cohabitation and marriage, using a multinomial logit model of current partnership type. The analysis is based on data from the 1999 Spanish Fertility Survey.

ABSTRACT The present study was carried out to evaluate the prevalence and hematological effects of Toxoplasma gondii in sheep and goat in district Dera Ghazi Khan. Blood samples (n=204) were collected comprise goats (n=101) and sheep (n=103) alongwith age, gender and breeds of animals. Samples were collected randomly from 25 flocks of 7 different union council Viz. Vehova, Tibbi Qaisrani, Lakhani, Kohar, Tuman Qaisrani, Nutkani and Kot Qaisrani of Tehsil Taunsa Sharif at least 4 animals from each flock. All ruminants divide into three groups based on age, breed and gender. The prevalence was detected through two different kits Viz. LAT and ELISA kit. The overall prevalence suspected in goats through LAT and ELISA kit was (35.64%), (32.67%) and in sheep was (25.24%), (23.30%) respectively. The Toxoplasma gondii had a significant effect on goats in age groups and non-significant all other groups of goats and sheep. Toxoplasma gondii had a significant effect on all hematological parameters like Hemoglobin, total leukocyte cells, granulocytes, lymphocytes, platelets, and red blood cells, except monocytes. In conclusion of the current study, toxoplasmosis is prevalent among ruminants, reveals the possibility of transmission to humans on the use of host animals as protein source.