Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera.

BACKGROUND: Endothelial cells (ECs) play a major role in malaria pathogenesis, as a point of direct contact of parasitized red blood cells to the blood vessel wall. The study of cytoskeleton structures of ECs, whose main functions are to maintain shape and provide strength to the EC membrane is important in determining the severe sequelae of Plasmodium falciparum malaria. The work investigated the cytoskeletal changes (microfilaments-actin, microtubules-tubulin and intermediate filaments-vimentin) in ECs induced by malaria sera (Plasmodium vivax, uncomplicated P. falciparum and complicated P. falciparum), in relation to the levels of pro-inflammatory cytokines. METHODS: Morphology and fluorescence intensity of EC cytoskeleton stimulated with malaria sera were evaluated using immunofluorescence technique. Levels of tumour necrosis factor (TNF) and interferon (IFN)-gamma (γ) were determined using enzyme-linked immunosorbent assay (ELISA). Control experimental groups included ECs incubated with media alone and non-malaria patient sera. Experimental groups consisted of ECs incubated with malaria sera from P. vivax, uncomplicated P. falciparum and complicated P. falciparum. Morphological scores of cytoskeletal alterations and fluorescence intensity were compared across each experiment group, and correlated with TNF and IFN-γ. RESULTS: The four morphological changes of cytoskeleton included (1) shrinkage of cytoskeleton and ECs with cortical condensation, (2) appearance of eccentric nuclei, (3) presence of "spiking pattern" of cytoskeleton and EC membrane, and (4) fragmentation and discontinuity of cytoskeleton and ECs. Significant damages were noted in actin filaments compared to tubulin and vimentin filaments in ECs stimulated with sera from complicated P. falciparum malaria. Morphological damages to cytoskeleton was positively correlated with fluorescence intensity and the levels of TNF and IFN-γ. CONCLUSIONS: ECs stimulated with sera from complicated P. falciparum malaria showed cytoskeletal alterations and increased in fluorescence intensity, which was associated with high levels of TNF and IFN-γ. Cytoskeletal changes of ECs incubated with complicated P. falciparum malaria sera can lead to EC junctional alteration and permeability changes, which is mediated through apoptotic pathway. The findings can serve as a basis to explore measures to strengthen EC cytoskeleton and alleviate severe malaria complications such as pulmonary oedema and cerebral malaria. In addition, immunofluorescence intensity of cytoskeleton could be investigated as potential prognostic indicator for malaria severity.

Potential Galactagogue Effect of Tri-Than-Thip Remedy on Milk Synthesis in Lactating Rats: Impact on Milk Production-Related Parameters.

Herbal galactagogues have been widely used as a treatment for postpartum hypogalactia due to the potential side effects associated with pharmacological therapy. Tri-Than-Thip (Tri-TT) is a Thai herbal medicine remedy that contains three main components: Cassia fistula, Pithecellobium dulce, and Ficus benjamina. These components are believed to have properties that contribute to milk production. However, despite the traditional use of Tri-TT, there is a lack of academic evidence supporting its efficacy in enhancing milk production. Therefore, the purpose of this study was to investigate the effect of Tri-TT on milk production and determine if it has a galactagogue effect. The weight suckle weight model was used to determine total milk production in lactating rats, while histological analysis was performed to assess the alveolar diameter of the mammary gland. The findings of this study revealed a significant increase in total milk production among lactating rats treated with 500 mg/kg of Tri-TT, compared to the control group. Furthermore, both the Tri-TT and Domperidone-treated groups exhibited a larger alveolar diameter of the mammary gland in comparison to the control group. In summary, these findings provide supportive evidence for the galactagogue activity of Tri-TT. The observed enhancement in milk production may be associated with Tri-TT could potentially be attributed to its ability to widen the alveolar diameter of the mammary gland, thereby facilitating increased milk volume.

Protective effects of Stephania pierrei tuber-derived oxocrebanine against LPS-induced acute lung injury in mice.

Acute lung injury and acute respiratory distress syndrome (ALI/ARDS) have high mortality rates. Though corticosteroids are commonly used for the treatment of these conditions, their efficacy has not been conclusively demonstrated and their use can induce various adverse reactions. Hence, the application of corticosteroids as therapeutic modalities for ALI/ARDS is limited. Meanwhile, the aporphine alkaloid oxocrebanine isolated from Stephania pierrei tubers has demonstrated anti-inflammatory efficacy in murine/human macrophage cell lines stimulated by lipopolysaccharide (LPS). Accordingly, the primary objectives of the present study are to investigate the anti-inflammatory effects of oxocrebanine on LPS-induced murine alveolar epithelial (MLE-12) cells and its efficacy against LPS-induced murine ALI. Results show that oxocrebanine downregulates the abundance of interleukin (IL)-1beta, IL-6, and inducible nitric oxide synthase, as well as the phosphorylation of nuclear factor-kappaB (NF-κB), stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), p38, protein kinase B (Akt), and glycogen synthase kinase-3beta signalling proteins in LPS-induced MLE-12 cells. Moreover, in a murine ALI model, oxocrebanine lowers lung injury scores and lung wet/dry weight ratios while reducing inflammatory cell infiltration. It also suppresses LPS-induced tumour necrosis factor-alpha and IL-6 in the bronchoalveolar lavage fluid and plasma. Moreover, oxocrebanine downregulates NF-κB, SAPK/JNK, p38, and Akt phosphorylation in the lung tissues of LPS-treated mice. Taken together, the foregoing results show that oxocrebanine provides significant protection against LPS-induced ALI in mice primarily by suppressing various inflammatory signalling pathways in alveolar epithelial cells and lung tissues. Hence, oxocrebanine might prove effective as an anti-inflammatory agent for the treatment of lung inflammation.

Profiles of CD4+, CD8+, and regulatory T cells and circulating cytokines in hookworm-infected children in southern Thailand.

Hookworm infection is the most common human helminthic infection in the rural areas of southern Thailand. There is little information on the induced cellular immune responses in hookworm-infected children. The present study aimed to investigate the cellular immune responses, regulatory T cells (Tregs), Th1-type cytokines (interleukin (IL)-2 and interferon (IFN)-γ), a Th2-type cytokine (IL-5) and IL-10, which is one of the cytokines secreted by Tregs in hookworm-infected children. Twenty-nine schoolchildren diagnosed with hookworm infections and 28 healthy controls were enrolled in the study. CD4+ and CD8+ T cells and Tregs in whole blood were analyzed using flow cytometry. Plasma IL-2, IL-5, IL-10 and IFN-γ concentrations were quantified by enzyme-linked immunosorbent assay (ELISA). The median CD4+ T cell frequency was significantly higher in hookworm-infected children than healthy controls. Compared to healthy controls, hookworm-infected children had a significantly increased absolute number of Tregs. No differences in circulating CD8+ T cell median frequency or absolute numbers were observed among hookworm-infected children or healthy controls. Elevated IL-2 and IL-10 concentrations were found in hookworm-infected children. Moreover, the absolute number of Tregs was significantly positively correlated with the plasma IL-10 concentration (rs = 0.406, P = 0.029). This study showed that hookworm-infected schoolchildren had significantly different immune responses than healthy controls, including an increase in the CD4+ T cell number, a significant induction of Tregs and significantly elevated circulating IL-10 levels. These alterations could be the mechanism underlying the immunomodulation that alleviates allergic diseases among hookworm-infected individuals.

Apoptotic changes and aquaporin-1 expression in the choroid plexus of cerebral malaria patients.

BACKGROUND: Cerebral malaria (CM) is associated with sequestration of parasitized red blood cells (PRBCs) in the capillaries. Often, the association of CM with cerebral oedema is related with high mortality rate. Morphological changes of the choroid plexus (CP) and caspase-3 expression in CM have not been reported. In addition, limited knowledge is known regarding the role of aquaporin (AQP)-1 in CM. The present study evaluated changes in the CP, explored apoptotic changes and AQP-1 expression in CP epithelial cells (CPECs) in fatal CM patients. METHODS: CP from fatal Plasmodium falciparum malaria patients (5 non-CM [NCM], 16 CM) were retrieved and prepared for histopathological evaluation. Caspase-3 and AQP-1 expressions in CPECs were investigated by immunohistochemistry. RESULTS: Histologically, apoptotic changes in CPECs were significantly observed in the CM group compared with the NCM and normal control (NC) groups (p < 0.05). These changes included cytoplasmic and nuclear condensation/shrinkage of CPECs and detachment of CPECs from the basement membrane. The apoptotic changes were positively correlated with caspase-3 expression in the nuclei of CPECs. In addition, AQP-1 expression in CPECs was significantly decreased in the CM group compared with the NCM and NC groups (all p < 0.001). A negative correlation (rs = - 0.450, p = 0.024) was documented between caspase-3 expression in the nuclei of CPECs and AQP-1. CONCLUSIONS: Apoptotic changes and altered AQP-1 expression may contribute to CPEC dysfunction and subsequently reduce cerebrospinal fluid production, affecting the water homeostasis in the brains of patients with CM.

Prevalence of soil-transmitted helminth infections and associated risk factors among elderly individuals living in rural areas of southern Thailand.

BACKGROUND: Soil-transmitted helminth (STH) infection is a neglected tropical disease affecting approximately 1.5 billion people worldwide. In past decades, most studies focused on STH infection in preschool-aged and school-aged children in different regions of Thailand. However, little is known about the prevalence and intensity of STH infection in the elderly population. Therefore, the aim of this study was to determine the current prevalence and intensity of STH infections and to identify associated risk factors among the elderly population. METHODS: A cross-sectional study was conducted from July to November 2019 to assess the prevalence of STH infections and associated risk factors among elderly populations living in five subdistricts of Thasala District, Nakhon Si Thammarat Province, Thailand. A total of 439 elderly individuals were selected using a random sampling technique. Each fresh stool sample was examined using the formalin ethyl acetate concentration technique (FECT), Kato-Katz thick smears and agar plate culture (APC). A structured questionnaire was used to obtain relevant information regarding associated risk factors for STH infection. RESULTS: The overall prevalence of STH infection was 15.7%. Hookworms (10.9%, 48/439) were the most prevalent STH species, followed by Strongyloides stercoralis (3.4%, 15/439) and Trichuris trichiura (2.1%, 9/439). Most elderly individuals infected with hookworms or T. trichiura had light-intensity infections. A higher prevalence of STH infection was observed among individuals aged older than 80 years (23.4%) than among those aged between 70 and 79 years (15.2%) and 60-69 years (14.5%). Males were 1.85-times more likely to present with STH infections than females. Not washing vegetables before eating increased the risk of STH infection by 3.19 times, while defecation in an open field increased the risk of STH infection by 2.65 times. CONCLUSIONS: The findings suggested that STH infection is prevalent, and that hookworms are the most common STH species among elderly populations in southern Thailand. Personal hygiene and deworming programs should be implemented among the elderly population to reduce the risk and prevent the spread of STH infections.

Release of endothelial activation markers in lungs of patients with malaria-associated acute respiratory distress syndrome.

BACKGROUND: Malaria-associated acute respiratory distress syndrome (MA-ARDS) is an understudied complication of malaria and is characterized by pulmonary inflammation and disruption of the alveolar-capillary membrane. Its pathogenesis remains poorly understood. Since endothelial activation plays an important role in other malarial complications, the expression of two endothelial activation markers, von Willebrand factor (VWF) and angiopoietin-2 (ANG-2), was investigated in the lungs of patients with MA-ARDS. METHODS: Post-mortem lung sections of Plasmodium falciparum-infected patients without alveolar oedema (NA), P. falciparum-infected patients with alveolar oedema (MA-ARDS), and uninfected people who died accidentally with no pathological changes to the lungs (CON) were immunohistochemically stained for VWF and ANG-2, and were evaluated with semi-quantitative analysis. RESULTS: Alveolar oedematous VWF and ANG-2 and intravascular VWF staining were significantly increased in patients with MA-ARDS versus infected and uninfected control groups. The levels of VWF in the alveolar septa and endothelial lining of large blood vessels of patients with MA-ARDS was significantly decreased compared to controls. ANG-2 expression was increased in the alveolar septa of malaria patients without alveolar oedema versus control patients, while ANG-2+ leukocytes were increased in the alveoli in both infected patient groups. CONCLUSIONS: This study documents a high level of VWF and ANG-2, two endothelial activation markers in the oedematous alveoli of post-mortem lung sections of Thai patients with MA-ARDS. Decreased detection of VWF in the endothelial lining of blood vessels, in parallel with an increased presence of intravascular VWF staining suggests marked endothelial activation and Weibel-Palade body release in the lungs of patients with MA-ARDS.

Prevalence of parasitic contamination of raw vegetables in Nakhon Si Thammarat province, southern Thailand.

BACKGROUND: Soil-transmitted helminth (STH) infections are major public health problems in poor and developing countries that require fecal contamination of the environment for transmission. The consumption of raw vegetables without proper washing is one of the main routes of intestinal parasite acquisition. Therefore, this study was designed to detect the prevalence of intestinal parasitic contamination in commonly consumed raw vegetables sold in three central open-air markets in Nakhon Si Thammarat province, southern Thailand. METHODS: A total of 265 fresh vegetable samples consisting of peppermint, lettuce, coriander, leek, gotu kola, celery, Chinese cabbage, culantro, Thai basil, and Chinese morning glory were purchased from three central open-air markets in the Mueang, Thasala and Sichon districts from December 2016 to March 2017. Each sample was washed with physiological saline, shaken for 15 min, and then allowed to sediment. Finally, sedimentation was performed via the sedimentation concentration technique and examined using light microscopy for the detection of pathogenic parasites. RESULTS: The overall prevalence of parasitic contamination was 35.1% (93/265). The most predominant parasite was hookworms (42.9%), followed by Strongyloides stercoralis (10.6%), Trichuris trichiura (2.6%), Ascaris lumbricoides (2.6%), and Toxocara spp. (2.6%). The highest level of contamination was found in celery, with a prevalence rate of 63.3% (19/30), while the lowest contamination level was found in Chinese morning glory, with a prevalence rate of 2.0% (2/30). The prevalence of intestinal parasite contamination in Mueang district (51.5%) was significantly higher than that in Thasala district (17.9%) and Sichon district (30.6%) (P < 0.001). CONCLUSION: The results of the present study demonstrate that consumption of vegetables with parasite contamination in this area represents a potential route for the transmission of parasitic infection, particularly hookworm infection. Therefore, it is necessary for health authorities to educate consumers about the proper washing of vegetables prior to consumption. Preventive methods such as wearing gloves and washing hands after handling vegetables should also be advocated to sellers who are at risk of acquiring STH infections via skin penetration.

Prevalence of malarial recurrence and hematological alteration following the initial drug regimen: a retrospective study in Western Thailand.

BACKGROUND: The hematological changes following the initial drug regimen has been poorly understood in Thailand. This study was designed to determine the prevalence of malaria parasite recurrence and hematological alteration of patients during the initial drug regimen. METHODS: A retrospective study was conducted at Phop Phra Hospital, Tak Province, located in northwestern Thailand. All data from patients who were diagnosed with Plasmodium spp. infection - including types of Plasmodium spp., clinical characteristics, and hematological parameters - were retrieved and analyzed. RESULTS: The results demonstrated that during years 2012-2018, 95 out of 971 patients (9.78%) were infected with malaria two or more times. The gender, nationality, symptom of headache, type of Plasmodium spp., and career of each patient were associated with recurrence (P-value< 0.05). Among patients treated with malarial drug, the leukocyte count and red cell distribution width (RDW) were significantly changed when compared to untreated patients with recurrence (P-value< 0.05). CONCLUSION: This study indicated the high prevalence of malarial recurrence in Tak Province, Western Thailand, and its relationship to certain characteristics of individuals. Patients who were treated with antimalarial drugs exhibited leukocyte and RDW changes following the initial drug regimen. This data could be useful for prompt detection, treatment, and prevention of malarial recurrence in endemic areas of Thailand.

Prevalence of intestinal parasitic infections and associated risk factors for hookworm infections among primary schoolchildren in rural areas of Nakhon Si Thammarat, southern Thailand.

BACKGROUND: Soil-transmitted helminth (STH) infections are among the most common type of infections worldwide and are widely distributed in tropical areas. In rural areas of southern Thailand where most land is used for agriculture, children are at risk of acquiring parasites, especially STHs. Assessing the current situation regarding parasitic infection in these areas is a prerequisite for developing appropriate control measures. This study is aimed at determining the prevalence of intestinal parasitic infections, the intensity of STH infections and the associated risk factors among primary schoolchildren in Nopphitam District, Nakhon Si Thammarat Province, Thailand. METHODS: A cross-sectional study involving 299 schoolchildren between 7 and 12 years of age was conducted between January and March 2016. A questionnaire administered by direct interviews was used to collect sociodemographic information and data on associated risk factors. Stool samples were processed using direct wet smears, formalin-ethyl acetate sedimentation concentration, and the modified Kato-Katz technique. RESULTS: The overall prevalence of intestinal parasites among the 299 children was 16% (48 of 299), with 32 children infected with hookworms (10.7%), 10 with Blastocystis hominis (3.3%), seven with Giardia intestinalis (1.6%), one with Enterobius vermicularis (0.3%), and one with Trichuris trichiura (0.3%). The hookworm infection intensity, measured by the median eggs per gram (EPG) of stool, was 1200 EPG (Interquartile range (IQR): 360-3200). Most children had light-intensity hookworm infections, but two had heavy-intensity infections. When participants included in the sample were classified by age, children 10-12 years old demonstrated higher intestinal parasite prevalence than those aged 7-9 years (adjusted odds ratio (AOR) = 2.3, 95% CI: 1.1-4.9, P = 0.030). Inadequate handwashing before meals was statistically associated with hookworm infections (AOR = 2.3; 95% CI: 1.1-4.8, P = 0.037). CONCLUSIONS: This study highlights that hookworms are the most prevalent STH infection in the study area. Older age group (10-12 years) and inadequate handwashing before meals were statistically associated with hookworm infections. Accordingly, appropriate strategies and education on personal and environmental hygiene should be implemented. Moreover, the cost-effectiveness of mass drug administration in this area should be further investigated.

Expression of cleaved caspase-3 in renal tubular cells in Plasmodium falciparum malaria patients.

AIM: In Plasmodium falciparum malaria, the clinical manifestation of acute kidney injury (AKI) is commonly associated with acute tubular necrosis (ATN) in the kidney tissues. Renal tubular cells often exhibit various degrees of cloudy swelling, cell degeneration, and frank necrosis. To study individual cell death, this study evaluates the degree of renal tubular necrosis in association with apoptosis in malarial kidneys. METHODS: Kidney tissues from P. falciparum malaria with AKI (10 cases), and without AKI (10 cases) were evaluated for tubular pathology. Normal kidney tissues from 10 cases served as controls. Tubular necrosis was assessed quantitatively in kidney tissues infected with P. falciparum malaria, based on histopathological evaluation. In addition, the occurrence of apoptosis was investigated using cleaved caspase-3 marker. Correlation between tubular necrosis and apoptosis was analyzed. RESULTS: Tubular necrosis was found to be highest in P. falciparum malaria patients with AKI (36.44% ± 3.21), compared to non-AKI (15.88% ± 1.63) and control groups (2.58% ± 0.39) (all p < 0.001). In the AKI group, the distal tubules showed a significantly higher degree of tubular necrosis than the proximal tubules (p = 0.021) and collecting tubules (p = 0.033). Tubular necrosis was significantly correlated with the level of serum creatinine (r = 0.596, p = 0.006), and the occurrence of apoptosis (r = 0.681, p = 0.001). CONCLUSION: In malarial AKI, the process of apoptosis occurs in ATN.

Prevalence of intestinal parasitic infection and associated risk factors among village health volunteers in rural communities of southern Thailand.

BACKGROUND: Intestinal parasitic infections remain prevalent and constitute a public health problem in certain rural areas of Thailand. Village health volunteers (VHVs), who are members of a Thai healthcare alliance, function as key providers of health prevention measures, disease control, and health education and share national health promotion campaigns with community members. This study is aimed at evaluating the prevalence, intensity, and risk factors for intestinal parasitic infection in VHVs in order to design community awareness and health education campaigns for the target population. METHODS: This cross-sectional study was conducted between January to April 2016 among village health volunteers (VHVs) from four sub-districts of Nopphitam District, Nakhon Si Thammarat Province, southern Thailand. Subjects for the study were selected using a simple random sampling method. Socio-demographic variables and risk factors were collected by a structured questionnaire. Stool specimens were collected and processed using direct wet mount and formol-ether concentration techniques to determine the presence of parasites and modified Kato-Katz thick smear to determine the intensity of infection. RESULTS: A total of 324 VHVs were enrolled. The overall prevalence of intestinal helminths was 9.3% (95% confidence interval [CI]: 6.3-13.0). The prevalence of hookworm, Strongyloides stercoralis, and Trichuris trichiura were 8.0% (95% CI: 5.3-11.5), 0.9% (95% CI: 0.2-2.7), and 0.3% (95% CI: 0-1.7), respectively. Mean intensity of hookworm infection was 1732 eggs per gram of stool. The prevalence was lower for protozoan infection than for helminth infection. Blastocystis hominis accounted for the highest percentage of intestinal protozoan infections 4.0% (95% CI: 2.2-6.8), followed by Giardia intestinalis 0.6% (95% CI: 0-2.2). No statistically significant difference was observed in the prevalence of intestinal parasitic infection among sub-districts (p > 0.05). Having dogs at home was associated with soil-transmitted helminth (STH) infection in study participants (Crude prevalence ratio [CPR]: 2.3; 95% CI: 1.0-5.2). CONCLUSIONS: This study is the first to describe the prevalence of intestinal parasitic infection VHVs from southern Thailand. Hookworm infection is more prevalent than other types of STH infection. The development of community awareness campaigns and appropriate control measures should be considered to reduce the prevalence of hookworm infection, especially among VHVs who are the key persons providing health education to the community.

Heme oxygenase-1 alleviates alcoholic liver steatosis: histopathological study.

Excessive alcohol consumption is one of the most important causes of hepatic steatosis, which involves oxidative stress. In particular, increased oxidative stress has been strongly linked to stimulation of the expression of heme oxygenase-1 (HO-1). This study aimed to investigate whether HO-1 could alleviates alcoholic steatosis in rats. Male Wistar rats were randomly divided into 4 groups: 1) the control group, 2) the EtOH group, 3) the EtOH + ZnPP-IX group and 4) the EtOH + Hemin group. Liver histopathology was investigated in weeks 1 and 4 after the start of the treatment period. Alcohol treatment significantly increased the hepatic malondialdehyde (MDA) levels, an oxidative stress marker. In addition, it increased the triglyceride, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in both weeks. Gross examination demonstrated a yellowish and slightly enlarged liver in the alcohol-treated rats. Hematoxylin and eosin (H&E) and Oil Red O staining indicated hepatic steatosis, which was characterized by diffuse, extensive fatty accumulation and discrete lipid droplets of variable size in hepatocytes of the alcohol-treated rats. Administration of the HO-1 inducer hemin resulted in upregulation of hepatic HO-1 gene expression, reduced the MDA, triglyceride, ALT and AST levels and alleviated alcoholic hepatic steatosis, whereas administration of the HO-1 inhibitor zinc protoporphyrin IX (ZnPP-IX) resulted in downregulation of hepatic HO-1 gene expression and could not alleviate alcoholic hepatic steatosis either week. In conclusion, HO-1 could alleviate alcoholic hepatic steatosis in male Wistar rats and may be useful in development of a new therapeutic approach.

Differential expression of matrix metalloproteinase-13 in association with invasion of breast cancer.

Matrix metalloproteinase-13 (MMP-13) has a potential role in tumour invasion and metastasis. However, its relevance to the prognosis of human breast cancer is poorly understood. The aim of this study is to investigate the expression patterns of MMP-13 protein and to determine its prognostic value in breast cancer, and to define its relation to the clinicopathological features. Immunohistochemistry analysis of MMP-13 was performed on formalin-fixed, paraffin-embedded sections of cancerous breast tissue (n = 76) and normal breast tissue (n = 20), all of which had clinicopathological information available. Based on the principle of immunoreactivity, the detection of MMP-13 on breast tissue was conducted using monoclonal antibodies against MMP-13. A semi-quantitative scoring system was used to assess the presence of, as well as the cellular localisation of MMP-13. MMP-13 expression was significantly greater in the cancerous breast tissues in comparison to those of normal breast tissues. In addition, high levels of MMP-13 expression were also found to be related to the positive detection of breast cancer cells in lymph nodes-amongst breast cancer patients. The results of this study showed that MMP-13 was frequently present in breast tumours, especially when tumours were accompanied by positive breast cancer cell detection in lymph nodes. This suggests that MMP-13 plays a potentially significant role in breast cancer invasion and metastasis.

Nuclear factor kappa B in urine sediment: a useful indicator to detect acute kidney injury in Plasmodium falciparum malaria.

BACKGROUND: Acute kidney injury (AKI) is one of the major complications of Plasmodium falciparum malaria, especially among non-immune adults. It has recently been revealed that activation of transcription factor nuclear factor kappa B (NF-κB) induces pro-inflammatory gene expression involved in the development of progressive renal inflammatory diseases. The aim of this study was to determine whether urinary sediment NF-κB p65 can act as a biomarker for AKI in patients with P. falciparum malaria. METHODS: Urinary sediments from malaria patients, including Plasmodium vivax malaria, uncomplicated P. falciparum malaria, complicated P. falciparum malaria without AKI (serum creatinine-Cr <3 mg/dl) and complicated P. falciparum malaria with AKI (Cr ≥3 mg/dl) were used to determine NF-κB p65 level by sandwich enzyme-linked immunosorbent assay (ELISA). Urinary sediments obtained from healthy controls were used as a normal baseline. Correlations between levels of urinary sediment NF-κB p65 and pertinent clinical data were analysed. RESULTS: Urinary sediment NF-κB p65 levels were significantly increased on the day of admission (day 0) and on day 7 post-treatment in complicated P. falciparum malaria patients with AKI, compared with those without AKI (p=0.001, p <0.001, respectively), P. vivax patients (all p <0.001) and healthy controls (all p <0.001). NF-κB p65 levels in urinary sediment cells showed a significant positive correlation with serum Cr (Day 0: rs=0.792; p <0.001, Day 7: rs=0.605; p <0.001) and blood urea nitrogen (BUN) (Day 0: rs=0.839; p <0.001, Day 7: rs=0.596; p <0.001). CONCLUSIONS: Urinary sediment NF-κB p65 level is a useful indicator for estimating renal tubular epithelial cell damage and subsequent development of AKI among patients with P. falciparum malaria.

Liver changes in severe Plasmodium falciparum malaria: histopathology, apoptosis and nuclear factor kappa B expression.

BACKGROUND: Liver involvement in severe Plasmodium falciparum infection is commonly a significant cause of morbidity and mortality among humans. The clinical presentation of jaundice often reflects a certain degree of liver damage. This study investigated the liver pathology of severe P. falciparum malaria as well as the regulation and occurrence of apoptosis in cellular components of formalin-fixed, paraffin-embedded liver tissues. METHODS: The liver tissues used in the study came from patients who died from P. falciparum malaria with hyperbilirubinaemia (total bilirubin (TB)≥ 51.3 µmol/L or 3 mg/dl) (12 cases), P. falciparum malaria without hyperbilirubinaemia (TB<51.3 µmol/L) (10 cases); and patients who died due to accidents, whose liver histology was normal (the control group) (10 cases). The histopathology of the liver tissue was studied by routine histology method. Caspase-3 and nuclear factor kappa B (NF-κB) p65 expressions were determined using immunohistochemistry. RESULTS: The severity of liver histopathology, occurrence of apoptosis and NF-κB p65 activation in P. falciparum malaria were associated with higher TB level. Significant correlations were found between NF-κB p65 expression and apoptosis in Kupffer cells and lymphocytes in the portal tracts. CONCLUSIONS: Hyperplastic Kupffer cells and portal tract inflammation are two main features found in the liver tissues of severe P. falciparum malaria cases. In addition, NF-κB is associated with Kupffer cells and lymphocyte apoptosis in severe P. falciparum malaria.

Glomerular changes and alterations of zonula occludens-1 in the kidneys of Plasmodium falciparum malaria patients.

BACKGROUND: The process of cytoadhesion in Plasmodium falciparum malaria infection causes signaling processes that lead to structural and functional changes at the cellular level. Histopathological changes of acute kidney injury (AKI) in P. falciparum malaria often involve glomerular proliferation, thickening of the glomerular basement membrane, acute tubular necrosis, and interstitial inflammation. Focusing on the glomeruli, this study aimed to investigate glomerular and tight junction-associated protein- zonula occludens-1 (ZO-1) changes in P. falciparum malaria patients. METHODS: Kidney tissues were grouped into P. falciparum with AKI (Cr ≥ 265 µmol/L or 3 mg/dl), P. falciparum without AKI (Cr < 265 µmol/L), and normal kidney tissues (control group). Glomerular cells and the glomerular area were quantified and compared in three experimental groups. The tight junction was investigated immunohistochemically using tight junction-associated protein, ZO-1, protein marker. A further immunofluorescence study was performed in an endothelial cell (EC)-parasitized red blood cell (PRBC) co-culture system, to evaluate the tight junction protein. RESULTS: Glomerular cell proliferation was significant in P. falciparum with AKI (Cr ≥ 265 µmol/L). By contrast, the glomerular area decreased significantly. ZO-1 expression was significantly decreased in the AKI group compared with normal kidneys, and in kidney tissues without AKI (p < 0.05). This was further confirmed by the depletion in ZO-1 localization in ECs co-cultured with PRBCs. CONCLUSIONS: In P. falciparum malaria with AKI, the decrease in glomerular area, despite glomerular cell proliferation, could be due to the collapse of cellular structures secondary to damaged tight junction-associated protein, ZO-1.

Hepatoprotective effect of curcumin and alpha-tocopherol against cisplatin-induced oxidative stress.

BACKGROUND: cis-Diammineplatinum (II) dichloride (cisplatin) is the important anti-cancer agent useful in treatment of various cancers. Unfortunately, it can produce unwanted side effects in various tissues, including the liver. The present study investigated the possible protective role of curcumin and α-tocopherol against oxidative stress-induced hepatotoxicity in rats upon cisplatin treatment. METHODS: Male Wistar rats were divided into five groups (n = 5). Saline and Cis groups, rats were intraperitoneal (i.p.) injected with normal saline and cisplatin [20 mg/kg body weight (b.w.)], respectively. Cis + α-tocopherol group, Cis + Cur group and Cis + α-tocopherol + Cur group, rats were pre-treated with a single dose of α-tocopherol (250 mg/kg b.w.), curcumin (200 mg/kg b.w.) and combined α-tocopherol with curcumin, respectively, for 24 h prior the administration of cisplatin. After 72 h of first injection, specimens were collected. Liver enzyme, lipid peroxidation biomarker, liver histopathology and gene expression of liver nicotinamide adenine dinucleotide phosphate (NADPH) oxidase were investigated. RESULTS: Cisplatin revealed a significant increase of hepatic malondialdehyde (MDA) levels and a significant reduction of hepatic superoxide dismutase (SOD) and catalase activities compared to the saline group. It elicited a marked increase of the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and demonstrated the liver pathologies including liver congestion, disorganization of hepatic cords and ground glass appearance of hepatocytes. It also demonstrated a significant increase of NADPH oxidase gene expression compared to saline group. Pre-treatment with combined curcumin and α-tocopherol improved the liver enzymes, lipid peroxidation biomarker, liver histopathology and gene expression of liver NADPH oxidase in cisplatin-treated rats. CONCLUSIONS: The findings indicate that pre-treatment with combined curcumin and α-tocopherol can protect cisplatin-induced hepatotoxicity including the biochemical, histological and molecular aspects. The down-regulations of NADPH oxidase gene expression may be involved in abrogating oxidative stress via reduction of reactive oxygen species (ROS) production.

Assessment of antimalarial activity of crude extract of Chan-Ta-Lee-La and Pra-Sa-Chan-Dang formulations and their plant ingredients for new drug candidates of malaria treatment: In vitro and in vivo experiments.

The emergence and spread of antimalarial drug resistance have become a significant problem worldwide. The search for natural products to develop novel antimalarial drugs is challenging. Therefore, this study aimed to assess the antimalarial and toxicological effects of Chan-Ta-Lee-La (CTLL) and Pra-Sa-Chan-Dang (PSCD) formulations and their plant ingredients. The crude extracts of CTLL and PSCD formulations and their plant ingredients were evaluated for in vitro antimalarial activity using Plasmodium lactate dehydrogenase enzyme and toxicity to Vero and HepG2 cells using the tetrazolium salt method. An extract from the CTLL and PSCD formulations exhibiting the highest selectivity index value was selected for further investigation using Peter's 4-day suppressive test, curative test, prophylactic test, and acute oral toxicity in mice. The phytochemical constituents were characterized using gas chromatography-mass spectrometry (GC-MS). Results showed that ethanolic extracts of CTLL and PSCD formulations possessed high antimalarial activity (half maximal inhibitory concentration = 4.88, and 4.19 g/mL, respectively) with low cytotoxicity. Ethanolic extracts of the CTLL and PSCD formulations demonstrated a significant dose-dependent decrease in parasitemia in mice. The ethanolic CTLL extract showed the greatest suppressive effect after 4 days of suppressive (89.80%) and curative (35.94%) testing at a dose of 600 mg/kg. Moreover, ethanolic PSCD extract showed the highest suppressive effect in the prophylactic test (65.82%) at a dose of 600 mg/kg. There was no acute toxicity in mice treated with ethanolic CTLL and PSCD extracts at 2,000 mg/kg bodyweight. GC-MS analysis revealed that the most abundant compounds in the ethanolic CTLL extract were linderol, isoborneol, eudesmol, linoleic acid, and oleic acid, whereas ethyl 4-methoxycinnamate was the most commonly found compound in the ethanolic PSCD extract, followed by 3-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-4H-chromen-4-one, flamenol, oleic acid amide, linoleic acid, and oleic acid. In conclusions, ethanolic CTLL and PSCD extracts exhibited high antimalarial efficacy in vitro. The ethanolic CTLL extract at a dose of 600 mg/kg exhibited the highest antimalarial activity in the 4-day suppressive and curative tests, whereas the ethanolic PSCD extract at a dose of 600 mg/kg showed the highest antimalarial activity in the prophylactic test.

Alpha-Glucosidase Inhibition, Antioxidant Activities, and Molecular Docking Study of Krom Luang Chumphon Khet Udomsak, a Thai Traditional Remedy.

Krom Luang Chumphon Khet Udomsak remedy (KKR) has traditionally been used as an alternative treatment, particularly for hyperglycemia; however, its therapeutic efficacy has not been scientifically validated. Thus, this study aims to investigate the potential inhibitory and antioxidant effects of α-glucosidase enzyme and characterize the chemical profile of KKR extracts using gas chromatography-mass spectrometry (GC-MS). The investigation highlights both KKR extracts as potent inhibitors of α-glucosidase, with the ethanolic extract of KKR (KKRE) displaying an IC50 value of 46.80 µg/mL and a noncompetitive mode of action. The combination of ethanolic and aqueous extracts of KKR (KKRE and KKRA, respectively) with acarbose exhibited a synergistic effect against the α-glucosidase. The KKRE extract displayed strong scavenging effects in the DPPH assay (IC50 156.3 µg/mL) and contained significant total phenolic (172.82 mg GAE/g extract) and flavonoid (77.41 mg QE/g extract) contents. The major component of KKRE is palmitic acid (15.67%). Molecular docking revealed that the major compounds interacted with key amino acid residues (ASP215, GLU277, HIS351, ASP352, and ARG442), which are crucial for inhibiting α-glucosidase. Notably, campesterin had a more significant influence on α-glucosidase than acarbose, with low binding energy. These findings underscore the significance of KKR in traditional medicine and suggest that it is promising treatment for diabetes mellitus. Further studies using animal model will provide valuable insights for advancing this research.