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The high mortality and morbidity rates of malaria remains to be a global problem, particularly in tropical regions. Cerebral malaria, the leading cause of death, possesses a complex pathogenesis involving host immune and parasite interactions. Lately, according to the gut-brain axis concept, the gut microbiota was identified to protect the host against systemic pathogen infection through the modulation of immune signaling pathways. In this study, we aimed to explore the effect of probiotic supplementation toward neurological outcomes of cerebral malaria in vivo. Twenty-eight male C57BL/6 mice were randomly divided into 5 groups after 2 weeks acclimatization period. All groups except the negative control were given oral probiotic supplementation with either Lactobacillus casei at 109 colony forming unit/100 µL (CFU/100 µL), Bifidobacterium longum at 109 CFU/100 µL, or the combination of each at 5x108 CFU/100 µL for five days before to six days after Plasmodium berghei infection. Neurological manifestations were recorded by using the SmithKline, Harwell, Imperial College, Royal Hospital, Phenotype Assessment (SHIRPA) score. The data was analyzed by using One-way ANOVA and post-hoc LSD. The results showed that there were significant differences of motor behavior (P < 0.01), muscle tone (P < 0.01), reflex and sensory (P < 0.05), and neuropsychiatric state (P < 0.05) between the control group and that receiving combination of L. casei and B. longum. The results suggested that probiotic supplementation showed promising effects in improving the neurological manifestations during cerebral malaria. Further studies are required to explicate the pathways involved in the immunological interplay. In the future, it is propitious that the mortalities by cerebral malaria could be mitigated through the optimal use of probiotics. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is one of the zoonotic protozoa parasites. It can prevalently infect humans and warmblooded animals, causing human health problems and substantial economic losses to the livestock industry worldwide. Chicken is one of the potential sources of toxoplasmosis, but there is no report of the prevalence of toxoplasmosis and their genotypes in free-range chickens in Libya. Aim: This study aims to conduct a survey of molecular prevalence and identify the T. gondii genotype in free-range chickens and its association with the risk factors of age, gender, and region in Northeastern Libya. Methods: This study was conducted by examining a total of 315 free-range chicken organs (brain and heart) derived from three administrative districts in Northeastern Libya. The molecular prevalence was determined by PCR technique using B1 gene amplification. and the T. gondii genotype was determined by nested PCR-RFLP of GRA6 gene amplicon with restriction enzymes (MseI). Results: The overall molecular prevalence of T. gondii in free-range chicken in all three districts was 9.5% (30/315), and the highest (15.4%) was in the Al-Marj district (p = 0.01; x² = 9.238). The highest prevalence of T. gondii by age was in chickens aged more than 2 years (p = 0.001; x² = 15.530). The difference in T. gondii prevalence in male and female chickens was not significant (p = 0.372; x² = 0.798). The predominant genotype I (93.3%) had identified at position 544 and 194 bp at the GRA6 marker, and only two positives were from genotype II (6.7%) at 700 and 100 bp fragments. Conclusion: The molecular prevalence of toxoplasmosis in free-range chicken in three districts in Northeastern Libya was 9.5%, and the highest rate was shown in the Al Marj district. Chicken by age more than 2 years had more risk to transmit toxoplasmosis in human. There was no different infection risk by consuming male or female free-range chicken. It is the first report to determine the predominant genotype, which was genotype I. [ABSTRACT FROM AUTHOR]

The phytochemical compounds contained in papaya leaves are known to have a galactopoietic effect. This study aims to analyze the effect of ethanol extract of Carica papaya leaves on β-casein gene expression, β- casein levels, total protein, and milk volume. This in vivo was an experimental study including a posttest control group that was conducted on one control group and three treatment groups. Each group consisted of six lactating rats. The control group rats were given ordinary food, while the treatment group rats, D1, D2, and D3, were given ethanol extract of Carica papaya leaves with the dose of 0.95 mg, 1.9 mg, and 3.8 mg/200 g Body weight (BW)/ day, respectively, from day 1 to day 13 of lactation. On day 14, all of the rats were sacrificed. Breastmilk volume taken from all breasts of lactating rats was measured individually in milliliters, β-casein gene expressions in the mammary tissues were measured using reverse transcription polymerase chain reaction (RT-PCR), while serum β-casein levels were measured using ELISA, and total protein was measured using bicinchoninic acid (BCA) protein assay. Statistical analysis was carried out using one-way ANOVA, Tukey test, and Games-Howell test at 95% confidence level. Milk volume, β-casein gene expression, β-casein levels, and total protein levels of all treatment rat groups were significantly higher than the control group (p<0.05). The increases of all parameters were consistent; the most effective dose was 1.9 mg/200g BW. Carica papaya leaf ethanol extract can increase milk volume, β-casein gene (Csn2) expression, β-casein levels, and total protein levels. [ABSTRACT FROM AUTHOR]

Background: Malaria screening for blood derived from any donors prior to transfusions is a standard procedure that should be performed; but, in fact, it is not routinely conducted. In case of the blood is infected with Plasmodium spp., the survival of parasites may be depending on, or even influencing, the profile of red blood cells (RBCs). Methods: This observational longitudinal study was conducted upon 55 bags of donor blood that randomly selected. Malaria infections were detected using Rapid Diagnostic Test/RDT with thin and thick blood smear confirmation. The changes of Plasmodium spp. viability and RBCs profiles, as well as other hematological parameters, were observed from the results of routine hematological examinations which were performed on days 1,7,14 and 21 of storage. Results: Among 55 blood samples, there were 17 and 38 bags, respectively, positive and negative for malaria, then used for analysis as the case and control groups. There were significant decreasing values (p< 0.05) of all routine blood examination parameters of donor blood, started from days 1, 7, 14, 21, and 28. There were no differences in decreasing profiles between those infected and non-infected donor blood (p> 0.05). On days 21 and 28 none of the positive samples still contained parasites. Conclusion: Erythrocytes profiles of donor blood significantly decreased with the duration of storage, but were not influenced by the presence of Plasmodium spp. [ABSTRACT FROM AUTHOR]

IL-17 in Toxoplasma gondii infection has an important role in miscarriage and low birth weight with many negative effects. This study explains the effect of hyperbaric oxygen (HBO2) administration on fetal body weight and IL-17 pregnant female rat levels. Rattus norvegicus were divided into four groups. IL-17 concentration was measured by serum ELISA. There was statistically significant reduction of IL-17 levels (79.42±30.36, 125.46±60.05, 201.17±70.33, 158.76±58.48, p = 0.001 for pregnant rats infected by Tachyzoite with HBO2 group, pregnant rats non-infected by Tachyzoit with HBO2 group, pregnant and infected without HBO2 group, and negative control, respectively). Although, there was no significant difference in the number of fetuses among groups (p = 0.096). The HBO2 treatment group had the heaviest of fetal weight among groups (0.98 [0.30 - 6.22] gram, p = 0.046). The results of the Spearman correlation test showed that only group B had a significant negative relationship between IL17 concentration and the mean fetal weight (p = 0.001). HBO2 therapy was found to be able to reduce IL-17 levels and result in rising of fetal weight in pregnant Rattus norvegicus and might prevent the occurrence of abortion. [ABSTRACT FROM AUTHOR]

Hormonal oestrus synchronization is often used to obtain the same gestational ages animal models, but it may not physiologic nor practical. This research was conducted to find out a simple method in obtaining physiologic pregnant experimental animals. We used 120 female rats and mice each, divided into 16 groups with 15 members each. They were classified according to the state of parities, pair or group mating, and synchronization of oestrous. The estrous synchronization is done by utilizing the natural phenomenon (Lee-Boot, Pheromone, and Whitten effect). All nulliparous and multiparous models then were simultaneously mated for one night in pairs (1:1) or in groups (1:5) subsequently. Pregnancy rates were noted based on the number of models delivering pups on day 19-23 post-mating. The highest pregnancy rate was noted in group of multiparous, synchronized and monogamous rats (86.7%), followed by nulliparous, synchronized and monogamous rats (73.3%) and multiparous, synchronized and monogamous mice (60%) (p < 0.05). Synchronization of the estrous and monogamous mating of rats or mice had significant roles in pregnancy rates (p < 0.05). To get a high number of pregnant rats or mice models, we suggest using the multiparous, estrous synchronized and monogamous simultaneous mating within one night. [ABSTRACT FROM AUTHOR]

Background. As a common ailment suffered by runners and athletes, Achilles tendinopathy pathogenesis is not well understood. To reveal its cellular and molecular changes, universal and standardized method to induce Achilles tendinopathy in animal model (in vivo) study should be established. There are 2 major guidelines to induce Achilles tendinopathy: run the rats on treadmill without inclination and run the rats on treadmill with 10° inclination. Objective. To compare available Achilles tendinopathy induction procedure hence determining the most effective procedure. Methods. Laboratory experimental animal study (in vivo) with post-test only control group approach. 2 procedures to induce Achilles tendinopathy on Wistar rats were performed: running at 15 m/min for 180 minutes, and running at 25 m/min for 15 minutes with 10° inclination for 60 minutes. Histological changes of Achilles tendon were converted into Bonar score while several inflammatory markers such as extracellular high mobility group box-1 (HMGB-1), tumor necrosis factor-alpha (TNF-α), C-X-C motif chemokine ligand-12 (CXCL-12), and macrophage-2 (M2) were measured. Results. Highest Bonar score was obtained from rats that run at 25 m/min for 15 minutes with 10° inclination for 60 minutes. Highest level of extracellular HMGB-1, TNF-α, and CXCL-12 was also obtained from similar group. This group also showed widest M2 coverage area. Conclusions. Running the rats at 25 m/min for 15 minutes with 10° inclination for 60 minutes is more effective than running the rats at 15 m/min for 180 minutes without inclination to induce Achilles tendinopathy. [ABSTRACT FROM AUTHOR]

Introduction: Toxoplasma gondii infection increases the production of Th1 cytokines that can cause fetal rejection. This study aims to determine the effect of hyperbaric oxygen therapy (HBOT) in the artery spiralis expressions of ICAM-1 in pregnant Rattus norvegicus infected by tachyzoite of Toxoplasma gondii. Material and Method: The rats were divided into four groups randomly. Determination of ICAM-1 expressions was performed on day-5 after HBOT and measured through Immunohistochemistry examination. Result: There was no significant difference in ICAM-1 expression among all groups (A and B [p <0.626], A and C [p <0.450], A and D [p <0.770], B and C (p <0.792), B and D [p <0.812], C and D [p <0.600]). However, the results showed that HBOT could not reduce ICAM-1 expression (6.4, 5.73, 5.4, 6.03, the mean ICAM-1 expression in HBO2 treatment group was the highest). Conclusion: This study concluded that HBOT could not reduce the expressions of ICAM-1 in artery spiralis with no significant difference among all groups. [ABSTRACT FROM AUTHOR]

Background: During pregnancy, the balanced dominance of the T helper17 response shifts to a Th2 response that is characterised by the production of IL-10, following the completion of the implantation process. Transforming growth factor-β (TGF-β) expression is associated with the completion of trophoblast invasion and placental growth. This study assessed the effect of malaria infection on the levels of IL-17, IL-10, and TGF-β in the plasma of pregnant mice with malaria. Methods: Seventeen pregnant BALB/C mice were divided into two groups: mice infected with Plasmodium berghei (treatment group) and uninfected mice (control group). The mice were sacrificed on day 18 post-mating. Parasitemia was measured by Giemsa staining. The levels of IL-17, IL-10, and TGF-β were measured by ELISA. Results: Using independent t test, the IL-17 levels in the treatment group were higher than those in the control group (P = 0.040). The IL-10 levels in the treatment group were lower than those in the control group (P = 0.00). There was no significant difference in the TGF-β levels (P = 0.055) between two groups. However, using SEM analysis the degree of parasitemia decreased the plasma TGF-β levels (tcount = 5.148; ≥ ttable= 1.96). SEM analysis showed that a high degree of parasitemia increased the IL-17 levels and decreased the IL-10 and TGF-β levels. Conclusion: Malaria infection during pregnancy interferes with the systemic balance by increasing the IL-17 levels and decreasing the IL-10 and TGF-β levels. [ABSTRACT FROM AUTHOR]

Background: Malaria in pregnancy leads to placental malaria. The primary pathogenesis of the complex fetal implications in placental malaria is tissue hypoxia due to sequestrations of Plasmodium falciparum -infected erythrocytes in the placenta. However, the pathomechanism of placental Plasmodium vivax infection has not been thoroughly investigated. Hypoxia-inducible factor-1α (HIF-1α) is a key transcriptional mediator of the response to hypoxic conditions, which interacts with the change and imbalances of many chemical mediators, including angiogenic factors, leading to fetal growth abnormality., Methods: This study was conducted cross-sectionally in Maumere, Sikka Regency, East Nusa Tenggara Province, previously known as one of the malaria endemic areas with a high incidence of low birth weight (LBW) cases. This study collected peripheral and umbilical blood samples and placental tissues from mothers who delivered their babies with LBW at the TC Hiller Regional Hospital. All of the blood samples were examined for parasites by microscopic and PCR techniques, while the plasma levels of VEGF, PlGF, VEGFR-1, VEGFR-2, and HIF-1α were determined using ELISA. The sequestration of infected erythrocytes and hemozoin was determined from placental histological slides, and the expression of placenta angiogenic factors was observed using the immunofluorescent technique., Results: In this study, 33 cases had complete data to be analyzed. Of them, 19 samples were diagnosed as vivax malaria and none of falciparum malaria. There were significant differences in Δ 10th percentile growth curve of baby's body weights and also all angiogenic factors in placental tissues {VEGF, PlGF, and VEGFR-1, VEGFR-2, and HIF-1α} between those infected and not infected cases (p<0.05), but not for VEGF and VEGFR-2 in the plasma., Conclusion: This study indicated that Plasmodium vivax sequestration may promote LBW through alterations and imbalances in angiogenic factors led by HIF-1α., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 Prasetyorini N et al.)

Background: During pregnancy, pregnant women are susceptible to malaria, contributing significantly to maternal and infant mortality., Objective: This research was conducted to study the effect of Plasmodium berghei infection in pregnant mice on fetal growth retardation through placental cell apoptosis and the change of local vascularization., Methods: Eighteen pregnant Balb/c strain mice resulting from simultanously mating were divided into two groups those were nine pregnant mice used as non infected group and nine pregnant mice infected with Plasmodium berghei on day 9th post mating used as infected group respectively. On day 15th of post mating, all of the pregnant mice were killed. Fetal weights were measured using analytic balance. Apoptosis of placental cells and VEGF expression in the placental tissue were measured using immunohistochemistry., Results: Result showed that there was sequestration of parasite-infected red blood cells (PRBCs) in intervillous space. Statistical analysis showed that the fetal weights in infected pregnant mice group was significantly lower than non infected one (p = 0.01), and the placental cell apoptosis in placental tissue of infected pregnant mice was significantly higher than the non infected one (p=0.00).There was also a significant difference on VEGF expression between infected group and non infected group (p= 0,00)., Conclusion: Plasmodium berghei infection in pregnant Balb/c mice can cause fetal growth retardation due to high of placental cell apoptosis and low VEGF expression., Competing Interests: There are no conflicts of interest., (© 2023 Kana Mardhiyyah, Tanto Hariyanto, Teguh Wahju Sardjono, Sri Winarsih, Tatit Nurseta, Loeki Enggar Fitri.)

Introduction: The saliva of mosquitoes has an important role in the transmission of several diseases, including malaria, and contains substances with vasomodulating and immunomodulating effects to counteract the host physiological mechanisms and enhance pathogen transmission. As immunomodulatory components, salivary gland proteins can induce the generation of specific IgG antibodies in the host, which can be used as specific biomarkers of exposure to Anopheles sundaicus . The objective of this study was to identify immunogenic proteins from the salivary glands of Anopheles sundaicus by reaction with sera from individuals living in malaria-endemic areas who are thus exposed to Anopheles mosquitoes., Methods: IgG antibodies targeting salivary gland proteins in serum samples from individuals living in malaria-endemic areas were measured by enzyme-linked immunosorbent assay (ELISA). Sera from healthy individuals living in non-endemic areas were used as negative controls. Determination of the presence of salivary gland immunogenic proteins was carried out by western blotting., Results: Sixteen bands appeared in sodium dodecyl sulfate polyacrylamide gel electrophoresis, with molecule weights ranging from 22 to 144kDa. Among the exposed individuals, IgG responses to salivary gland proteins were variable. Protein bands with molecular weights of 46, 41, 33, and 31kDa were the most immunogenic. These immunogenic proteins were consistently recognized by pooled serum and individual samples from people living in malaria-endemic areas but not by negative controls., Conclusions: These results support the potential use of immunogenic proteins from the salivary glands of Anopheles as candidate markers of bite exposure or in malaria vaccines.

Background: During pregnancy, the balanced dominance of the T helper17 response shifts to a Th2 response that is characterised by the production of IL-10, following the completion of the implantation process. Transforming growth factor-β (TGF-β) expression is associated with the completion of trophoblast invasion and placental growth. This study assessed the effect of malaria infection on the levels of IL-17, IL-10, and TGF-β in the plasma of pregnant mice with malaria., Methods: Seventeen pregnant BALB/C mice were divided into two groups: mice infected with Plasmodium berghei (treatment group) and uninfected mice (control group). The mice were sacrificed on day 18 post-mating. Parasitemia was measured by Giemsa staining. The levels of IL-17, IL-10, and TGF-β were measured by ELISA., Results: Using independent t test, the IL-17 levels in the treatment group were higher than those in the control group (= = 0.040). The IL-10 levels in the treatment group were lower than those in the control group (= = 0.00). There was no significant difference in the TGF-β levels (= = 0.055) between two groups. However, using SEM analysis the degree of parasitemia decreased the plasma TGF-β levels (tcount = 5.148; ≥ ttable = 1.96). SEM analysis showed that a high degree of parasitemia increased the IL-17 levels and decreased the IL-10 and TGF-β levels., Conclusion: Malaria infection during pregnancy interferes with the systemic balance by increasing the IL-17 levels and decreasing the IL-10 and TGF-β levels.