Molecular detection and species identification of Plasmodium spp. infection among imported cases in Eastern China, 2012-2018.

Due to the increasing number of returnees from malaria endemic areas, imported malaria has become a public health challenge in China. To better understand the characteristics of imported Plasmodium species and adjust appropriate strategies for malaria prevention and control in Eastern China, we conducted molecular detection and species identification on 1282 imported malaria cases in Shandong Province between 2012 and 2018. The findings showed that P. falciparum was predominant, particularly in cases imported from Africa. P. vivax was the dominant species imported from Asian countries. Additionally, imported P. ovale and P. malariae emerged in the province. Further surveillance and control of imported malaria among returnees from Africa and Southeast Asia is needed to be strengthened in Eastern China.

Preparation of Neohesperidin-Taro Starch Complex as a Novel Approach to Modulate the Physicochemical Properties, Structure and In Vitro Digestibility.

Neohesperidin (NH), a natural flavonoid, exerts multiple actions, such as antioxidant, antiviral, antiallergic, vasoprotective, anticarcinogenic and anti-inflammatory effects, as well as inhibition of tumor progression. In this study, the NH-taro starch complex is prepared, and the effects of NH complexation on the physicochemical properties, structure and in vitro digestibility of taro starch (TS) are investigated. Results showed that NH complexation significantly affected starch gelatinization temperatures and reduced its enthalpy value (ΔH). The addition of NH increased the viscosity and thickening of taro starch, facilitating shearing and thinning. NH binds to TS via hydrogen bonds and promotes the formation of certain crystalline regions in taro starch. SEM images revealed that the surface of NH-TS complexes became looser with the increasing addition of NH. The digestibility results demonstrated that the increase in NH (from 0.1% to 1.1%, weight based on starch) could raise RS (resistant starch) from 21.66% to 27.75% and reduce RDS (rapidly digestible starch) from 33.51% to 26.76% in taro starch. Our work provided a theoretical reference for the NH-taro starch complex's modification of physicochemical properties and in vitro digestibility with potential in food and non-food applications.

Molecular surveillance of artemisinin resistance-related Pfk13 and pfcrt polymorphisms in imported Plasmodium falciparum isolates reported in eastern China from 2015 to 2019.

BACKGROUND: Artemisinin-based combination therapy (ACT) has been recommended as the first-line treatment by the World Health Organization to treat uncomplicated Plasmodium falciparum malaria. However, the emergence and spread of P. falciparum resistant to artemisinins and their partner drugs is a significant risk for the global effort to reduce disease burden facing the world. Currently, dihydroartemisinin-piperaquine (DHA-PPQ) is the most common drug used to treat P. falciparum, but little evidence about the resistance status targeting DHA (ACT drug) and its partner drug (PPQ) has been reported in Shandong Province, China. METHODS: A retrospective study was conducted to explore the prevalence and spatial distribution of Pfk13 and Pfcrt polymorphisms (sites of 72-76, and 93-356) among imported P. falciparum isolates between years 2015-2019 in Shandong Province in eastern China. Individual epidemiological information was collected from a web-based reporting system were reviewed and analysed. RESULTS: A total of 425 P. falciparum blood samples in 2015-2019 were included and 7.3% (31/425) carried Pfk13 mutations. Out of the isolates that carried Pfk13 mutations, 54.8% (17/31) were nonsynonymous polymorphisms. The mutant alleles A578S, Q613H, C469C, and S549S in Pfk13 were the more frequently detected allele, the mutation rate was the same as 9.7% (3/31). Another allele Pfk13 C580Y, closely associated with artemisinin (ART) resistance, was found as 3.2% (2/31), which was found in Cambodia. A total of 14 mutant isolates were identified in Western Africa countries (45.2%, 14/31). For the Pfcrt gene, the mutation rate was 18.1% (77/425). T76T356 and T76 were more frequent in all 13 different haplotypes with 26.0% (20/77) and 23.4% (18/77). The CVIET and CVIKT mutant at loci 72-76 have exhibited a prevalence of 19.5% (15/77) and 3.9% (3/77), respectively. The CVIET was mainly observed in samples from Congo (26.7%, 4/15) and Mozambique (26.7%, 4/15). No mutations were found at loci 97, 101 and 145. For polymorphisms at locus 356, a total of 24 isolates were identified and mainly from Congo (29.2%, 7/24). CONCLUSION: These findings indicate a low prevalence of Pfk13 in the African isolates. However, the emergence and increase in the new alleles Pfcrt I356T, reveals a potential risk of drug pressure in PPQ among migrant workers returned from Africa. Therefore, continuous molecular surveillance of Pfcrt mutations and in vitro susceptibility tests related to PPQ are necessary.

High genetic diversity in Plasmodium falciparum isolates among Chinese migrant workers returnee from Africa.

Malaria is one of the most important parasitic diseases that causes a serious public health problem. The genetic diversity of malaria parasites may affect malaria transmission and malaria control strategies. In China, imported malaria was significantly increased in recent years, among which numerous migrant workers were infected with Plasmodium falciparum from Africa. However, little was known about genetic diversity of these populations in China. In this study, we evaluated genetic polymorphism and allele frequencies of msp1, msp2, and glurp genes in P. falciparum among Chinese migrant workers returnee from Africa between 2013 and 2017. Of the 381 P. falciparum isolates, 89.0% for msp1 gene, 71.7% for msp2 gene, and 78.0% for glurp gene were successfully genotyped. In msp1, 29 different alleles were observed, among which the K1 allelic family (71.7%) was predominant. In msp2, 21 different alleles were detected, of which the 3D7 allelic family (91.2%) was more frequent than FC27 allelic family (72.5%). For glurp, 12 individual alleles were detected in the samples. Taken together, the findings showed a high genetic diversity of these isolates, which provided the baseline data for African P. falciparum population imported to China.

Characterization of pfmdr1, pfcrt, pfK13, pfubp1, and pfap2mu in Travelers Returning from Africa with Plasmodium falciparum Infections Reported in China from 2014 to 2018.

The artemisinin-based combination therapies (ACTs) used to treat Plasmodium falciparum in Africa are threatened by the emergence of parasites in Asia that carry variants of the Kelch 13 (K13) locus with delayed clearance in response to ACTs. Single nucleotide polymorphisms (SNPs) in other molecular markers, such as ap2mu and ubp1, were associated with artemisinin resistance in rodent malaria and clinical failure in African malaria patients. Here, we characterized the polymorphisms in pfmdr1, pfcrt, pfK13, pfubp1, and pfap2mu among African isolates reported in Shandong and Guangxi provinces in China. Among 144 patients with P. falciparum returning from Africa from 2014 to 2018, pfmdr1 N86Y (8.3%) and pfcrt K76T (2.1%) were the major mutant alleles. The most common genotype for pfcrt was I74E75T76 (8.3%), followed by E75T76 (2.1%). For K13 polymorphisms, a limited number of mutated alleles were observed, and A578S was the most frequently detected allele in 3 isolates (2.1%). A total of 27.1% (20/144) of the isolates were found to contain pfubp1 mutations, including 6 nonsynonymous and 2 synonymous mutations. The pfubp1 genotypes associated with artemisinin resistance were D1525E (10.4%) and E1528D (8.3%). Furthermore, 11 SNPs were identified in pfap2mu, and S160N was the major polymorphism (4.2%). Additionally, 4 different types of insertions were found in pfap2mu, and the codon AAT, encoding aspartic acid, was more frequently observed at codons 226 (18.8%) and 326 (10.7%). Moreover, 4 different types of insertions were observed in pfubp1 at codon 1520, which was the most common (6.3%). These findings indicate a certain degree of variation in other potential molecular markers, such as pfubp1 and pfap2mu, and their roles in either the parasite's mechanism of resistance or the mode of action should be evaluated or elucidated further.

Prevalence of Plasmodium falciparum Kelch 13 (PfK13) and Ubiquitin-Specific Protease 1 (pfubp1) Gene Polymorphisms in Returning Travelers from Africa Reported in Eastern China.

Delayed clearance of Plasmodium falciparum by artemisinin-based combination therapies (ACTs) has already been observed for African isolates. Here, we aimed to investigate the prevalence, among travelers returning from African countries, of polymorphisms in two genes correlated with delayed parasite clearance (encoding P. falciparum Kelch 13 [PfK13] and ubiquitin-specific protease 1 [pfubp1]) reported in eastern China and to provide baseline data for antimalarial drug resistance (ART) surveillance and evaluation. A total of 153 filter paper blood spots collected in 2017-2019 from patients with uncomplicated P. falciparum cases in Anhui and Shandong Provinces were included in this study. Among them, 3.3% (5/153) of the isolates carried PfK13 mutations, and 3 of them harbored the same synonymous mutation, C469C. A total of 13.1% (20/153) of the isolates were found to contain pfubp1 mutations, and all were nonsynonymous. The pfubp1 genotypes associated with ART that occurred in this study included E1528D (6.5% [10/153]) and D1525E (2.6% [4/153]). However, a high prevalence of the previously unreported mutation E1531D (5.9% [9/153]) was also detected. In addition, two types of deletions (encoding KID and KIE, respectively) and two types of insertions (encoding KYE and KYDKYD, respectively) were found in 16 isolates and 6 isolates, respectively. This study showed limited variation in PfK13 among travelers returning from African countries and suggested other potential molecular markers, such as pfubp1, for use in the surveillance of African isolates in ACT susceptibility studies. Further clinical trial research is under way to investigate these PfK13 and pfubp1 mutations, as well as other candidate molecular markers, and their roles in delaying parasite clearance.

Effect of Potato Pulp Pectic Polysaccharide on the Stability of Acidified Milk Drinks.

In order to broaden the application of potato pulp pectic polysaccharide (PPP) in stabilizing acidified milk drinks (AMDs) and investigate the stabilizing effect and physical properties of AMDs prepared with PPP, a comparative study was made among PPP, commercial high methoxyl pectin (HMP) and low methoxyl pectin (LMP). The zeta potential, rheology, particle size and serum separation of AMDs were evaluated after preparing with PPP, HMP and LMP, respectively. Results indicated that PPP led to lower serum separation than LMP (14.65% for AMDs prepared with 0.5% PPP compared to 25.05% for AMDs prepared with 0.5% LMP), but still higher than HMP (9.09% for AMDs prepared with 0.5% HMP). However, narrower particle size distribution and lower viscosity of AMDs was achieved by PPP than by LMP and HMP. PPP can electrostatically adsorb on the surface of casein and its abundant neutral sugar side chains would provide steric hindrance to prevent casein flocculation in AMDs. Our results might provide some new ideas for the application of PPP in improving the stability of AMDs.

Physicochemical and Digestion Properties of Potato Starch Were Modified by Complexing with Grape Seed Proanthocyanidins.

Dietary intake of potato starch could induce a dramatic increase in blood glucose and is positively associated with chronic metabolic diseases (type II diabetes, cardiovascular disease, etc.). Grape seed proanthocyanidins (GSP) are known to decrease starch digestion by inhibiting digestive enzymes or changing the physicochemical properties of starch. In the present study, GSP were complexed with potato starch to prepare polyphenol-starch complexes. The physiochemical properties and digestibility of complexes were investigated by in vitro digestion model, X-ray diffraction, differential scanning calorimetry, rapid visco analyzer, Fourier transform infrared spectroscopy as well as texture profile analysis. Results indicated that the peak viscosity, breakdown, trough, and setback of the complexes disappeared, replaced by a special continuous increase in paste viscosity. The complexes showed a lower final viscosity and higher thermal stability with the increasing binding amount of GSP. GSP decreased the hardness of the complexes' gel significantly. FT-IR indicated that GSP might interact with potato starch through noncovalent forces. Additionally, the complexes also showed a higher content of slowly digestible starch and resistant starch than that of the native starch. Thus, we inferred that the addition of GSP could modify the digestibility of potato starch and be an optional way to modify the starch with lower digestion.

Mutation Profile of pfdhfr and pfdhps in Plasmodium falciparum among Returned Chinese Migrant Workers from Africa.

We evaluated markers of sulfadoxine-pyrimethamine (SP) resistance in Plasmodium falciparum among 254 returned migrant workers in China from Africa from 2013 to 2016. High prevalences of pfdhfr (97.2%) and pfdhps (96.5%) mutations were observed. The partially resistant genotype was homogeneously distributed in Africa with a modestly high prevalence (48%), whereas the super resistant genotype was only found in West Africa with a very low frequency (1.2%). The findings provided baseline data about the molecular markers of SP resistance.

Critical roles of soluble starch synthase SSIIIa and granule-bound starch synthase Waxy in synthesizing resistant starch in rice.

Changes in human lifestyle and food consumption have resulted in a large increase in the incidence of type-2 diabetes, obesity, and colon disease, especially in Asia. These conditions are a growing threat to human health, but consumption of foods high in resistant starch (RS) can potentially reduce their incidence. Strategies to increase RS in rice are limited by a lack of knowledge of its molecular basis. Through map-based cloning of a RS locus in indica rice, we have identified a defective soluble starch synthase gene (SSIIIa) responsible for RS production and further showed that RS production is dependent on the high expression of the Waxya (Wxa ) allele, which is prevalent in indica varieties. The resulting RS has modified granule structure; high amylose, lipid, and amylose-lipid complex; and altered physicochemical properties. This discovery provides an opportunity to increase RS content of cooked rice, especially in the indica varieties, which predominates in southern Asia.

Malaria control and prevention towards elimination: data from an eleven-year surveillance in Shandong Province, China.

BACKGROUND: Shandong Province experienced a declining malaria trend of local-acquired transmission, but the increasing imported malaria remains a challenge. Therefore, understanding the epidemiological characteristics of malaria and the control and elimination strategy and interventions is needed for better planning to achieve the overall elimination goal in Shandong Province. METHODS: A retrospective study was conducted and all individual cases from a web-based reporting system were reviewed and analysed to explore malaria-endemic characteristics in Shandong from 2005 to 2015. Annual malaria incidence reported in 2005-2015 were geo-coded and matched to the county-level. Spatial cluster analysis was performed to evaluate any identified spatial disease clusters for statistical significance. The space-time cluster was detected with high rates through the retrospective space-time analysis scanning using the discrete Poisson model. RESULTS: The overall malaria incidence decreased to a low level during 2005-2015. In total, 1564 confirmed malaria cases were reported, 27.1% of which (n = 424) were indigenous cases. Most of the indigenous case (n = 339, 80.0%) occurred from June to October. However, the number and scale of imported cases have been increased but no significant difference was observed during months. Shandong is endemic for both Plasmodium vivax (n = 730) and Plasmodium falciparum (n = 674). The disease is mainly distributed in Southern (n = 710) and Eastern region (n = 424) of Shandong, such as Jinning (n = 214 [13.7%]), Weihai (n = 151 [9.7%]), and Yantai (n = 107 [6.8%]). Furthermore, the spatial cluster analysis of malaria cases from 2005 to 2015 indicated that the diseased was not randomly distributed. For indigenous cases, a total of 15 and 2 high-risk counties were determined from 2005 to 2009 (control phase) and from 2010 to 2015 (elimination phase), respectively. For imported cases, a total of 26 and 29 high-risk counties were determined from 2005 to 2009 (control phase) and from 2010 to 2015 (elimination phase), respectively. The method of spatial scan statistics identified different 13 significant spatial clusters between 2005 and 2015. The space-time clustering analysis determined that the most likely cluster included 14 and 19 counties for indigenous and imported, respectively. CONCLUSIONS: In order to cope with the requirements of malaria elimination phase, the surveillance system should be strengthened particularity on the frequent migration regions as well as the effective multisectoral cooperation and coordination mechanisms. Specific response packages should be tailored among different types of cities and capacity building should also be improved mainly focus on the emergence response and case management. Fund guarantees for scientific research should be maintained both during the elimination and post-elimination phase to consolidate the achievements of malaria elimination.

Characteristics of Imported Malaria and Species of Plasmodium Involved in Shandong Province, China (2012-2014).

Malaria remains a serious public health problem in Shandong Province, China; therefore, it is important to explore the characteristics of the current malaria prevalence situation in the province. In this study, data of malaria cases reported in Shandong during 2012-2014 were analyzed, and Plasmodium species were confirmed by smear microscopy and nested-PCR. A total of 374 malaria cases were reported, 80.8% of which were reported from 6 prefectures. Of all cases, P. falciparum was dominant (81.3%), followed by P. vivax (11.8%); P. ovale and P. malariae together accounted for 6.4% of cases. Notably, for the first time since 2012, no indigenous case had been reported in Shandong Province, a situation that continued through 2014. Total 95.2% of cases were imported from Africa. The ratio of male/female was 92.5:1, and 96.8% of cases occurred in people 20-54 years of age. Farmers or laborers represented 77.5% of cases. No significant trends of monthly pattern were found in the reported cases. All patients were in good condition after treatment, except for 3 who died. These results indicate that imported malaria has increased significantly since 2012 in Shandong Province, especially for P. falciparum, and there is an emergence of species diversity.

[The Effects of NF-kappaB Signalling on the TNF-α induced Ratios of MMPs to TIMPs in Chondrocytes].

OBJECTIVES: To explore the influence of the NF-κB inhibitor (bay11-7082) on tumor necrosis factor-α (TNF-α)-induced different ratios of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in chondrocytes. METHODS: Chondrocytes were isolated from the knee joint of a 1-day old mouse by trypsin digestion method. Hematoxylin-Eosin (HE) stain was used to show the morphology of isolated chondrocytes; Semi-quantitative PCR was applied to analyze the influence of bay11-7082 on gene expressions of TNF-α-induced MMPsand TIMPsin chondrocytes; Zymography was used to elucidate activities of the gelatinases induced by TNF-α and/or bay11-7082. RESULTS: TNF-α up-regulated gene expressiosn of the MMPsand TIMPs(P<0.05). The ratios of MMPs/TIMPswere mostly increased except the part of MMP-1. Bay11-7082 could reduce TNF-α-induced MMPsand TIMPsgene expressions, and could make the increased ratio of MMPs/TIMPsdropped to the normal level of chondrocytes. Similar results were observed at the protein level of the gelatinases by zymography. CONCLUSIONS: TNF-α-induced high ratios of MMPs/TIMPs could partiallyexplain over-degradation of cartilage extracellular matrix in osteoarthritis (OA). Blockage of NF-κB with bay11-7082 might provide a possible therapeutic strategy for the OA deterioration.

[Detection of Genetic Mutations Associated with Drug Resistance in Imported Plasmodium falciparum].

Objective: To investigate the mutation of genes associated with drug resistance (Pfcrt, Pfmdr1, Pfdhfr and K13） in imported Plasmodium falciparum in Shandong Province. Methods: Blood was collected from 94 falciparum malaria cases who returned from Africa in 2014. Genomic DNA for P. falciparum was extracted from the blood samples and nested PCR was performed using primers specifically designed for Pfcrt, Pfmdr1, Pfdhfr and K13. The PCR products were sequenced. Gene mutations were analyzed by sequence alignment. Results: The 94 imported cases were from 18 African countries. Nested PCR was successful on DNA from all the blood samples except for Pfcrt amplification in one sample. Sequence analysis revealed three types of mutations Pfcrt K76T （36.6%, 34/93）, Pfmdr1 N86Y （21.3%, 20/94）, and Pfdhfr S108N （98.9%, 93/94） （χ2=127.5， P<0.05）. K13 C580Y mutation was not found. Co-occurrence of K76T, N86Y, and S108N was found in 6 blood samples （6.5%）, which were imported from Liberia（2）, Angola（1）, Equatorial guinea（1）, Congo（1）, and Guinea（1）. Co-occurrence of K76T and S108N mutations was found in 28 samples（30.1%）, and that of N86Y and S108N in 14 samples （15.1%）. Forty-four samples（47.3%） harbored S108N mutation only, and one sample was null for any of the mutations. Conclusion: There are mutations in Pfcrt, Pfmdr1, and Pfdhfr in imported Plasmodium falciparum in Shandong Province. No mutation was found for the K13 gene.

Physicochemical and crystalline properties of heat-moisture-treated rice starch: combined effects of moisture and duration of heating.

BACKGROUND: Currently there is much interest in the application of physical modification techniques such as heat-moisture treatment (HMT). The effects of HMT on normal and waxy rice starches, subject to different levels of moisture content and duration of heating, were investigated. RESULTS: Water solubility index (determined at 90 °C) decreased after HMT for normal and waxy rice starches, while swelling power (determined at 90 °C) showed inconsistent results (decrease for normal type, increase for waxy type) after HMT. Values in pasting parameters of normal and waxy rice starch increased initially, but the extent of increase slowed down with moisture content and length of treatment increasing. HMT decreased gelatinization temperatures with 4 h and 8 h treatment, but when length of treatment was prolonged to 16 h gelatinization temperature increased. Degree of crystallinity decreased for all treatments, and decreased much more at higher levels of moisture content. CONCLUSION: Variations in levels of moisture content and duration of heating had significant effects on physicochemical and crystalline properties to different extents.

TCAB1: a potential target for diagnosis and therapy of head and neck carcinomas.

BACKGROUND: WRAP53, including α, ß and γ isoforms, plays an important role not only in the stability of p53 mRNA, but also in the assembly and trafficking of the telomerase holoenzyme. It has been considered an oncogene and is thought to promote the survival of cancer cells. The aim of this study was to detect the role of TCAB1 (except WRAP53α) in the occurrence and development of head and neck carcinomas. METHODS: Immunohistochemistry was used to detect the TCAB1 expression in clinical specimen sections and performed western blotting to check the TCAB1 expression levels in cell lines. TCAB1 was depleted using shRNA lentivirus and the knockdown efficiency was assessed using q-PCR and Western blotting. We performed CCK-8 assays and flow cytometry to check the cell proliferation potential and used the trans-well assay to test the invasion ability in vitro. Xenografts were used to detect the tumor formation potential in vivo. Moreover, we performed cDNA microarray to investigate the candidate factors involved in this process. RESULTS: We observed a notable overexpression of TCAB1 in head and neck carcinoma clinical specimens as well as in carcinoma cell lines. Knockdown of TCAB1 decreased the cellular proliferation potential and invasion ability in vitro. cDNA microarray analysis suggested the possible involvement of several pathways and factors associated with tumorigenesis and carcinoma development in the TCAB1-mediated regulation of cancers. Furthermore, the xenograft assay confirmed that the depletion of TCAB1 would inhibit tumor formation in nude mice. The immunohistochemistry results of the mice tumor tissue sections revealed that the cells in shTCAB1 xenografts showed decreased proliferation potential and increased apoptotic trend, meanwhile, the angiogenesis was inhibited in the smaller tumors form shTCAB1 cells. CONCLUSIONS: Our study demonstrated that depletion of TCAB1 decreased cellular proliferation and invasion potential both in vitro and in vivo. The data indicated that TCAB1 might facilitate the occurrence and development of head and neck carcinomas. In future, TCAB1 might be useful as a prognostic biomarker or a potential target for the diagnosis and therapy of head and neck carcinomas.

Isolation and characterization of natural nano starch from amaranth starch.

Nano starch exhibits many advantages for application in diverse fields. Amaranth starch consisted of starch particle aggregates, isolated amaranth starch, and few natural nano starch (NNS), while NNS (0.92 ± 0.12 µm) was successfully isolated for the first time. Compared with the isolated amaranth starch, NNS showed smaller particle size but larger molecular weight, suggesting that the molecules arranged densely. NNS had a weak A-type crystal structure because of its more content of short starch chains, but higher amylose content resulted in the increase of its gelatinization temperature. The special NNS, owning several different physicochemical properties from amaranth starch, can open new ways for the production and application of nano biomass materials.

Effect of hawthorn polyphenol extracts on the physicochemical properties and digestibility of corn starch.

To investigate the effect of hawthorn polyphenols on the physicochemical properties and digestibility of corn starch, different proportions (1%, 2%, 3%, and 4% [w/w]) of hawthorn polyphenol extracts (HPEs) were mixed with corn starch, and their physicochemical properties and digestive properties were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, Rapid Visco Analysis, differential scanning calorimetry, and in vitro/in vivo analysis. Results indicated that small V-type crystal starch tended to be formed in the samples, and the addition of HPEs reduced the viscosity, prolonged the gelatinization temperature of corn starch, and increased the proportion of slowly digestible starch and resistant starch of the corn starch, which accounted for 36.32% ± 1.05% and 33.32% ± 4.07%, respectively. Compared with the raw corn starch, the postprandial blood glucose of mice that were administered the hawthorn polyphenols decreased significantly: the blood glucose peak (30 min) decreased from 14.30 ± 1.52 to 11.77 ± 1.21 mmol/L. Our study might provide some basic theoretical support for the application of hawthorn polyphenols in healthy starchy food processing.

Disruption of the blood-brain barrier after generalized tonic-clonic seizures correlates with cerebrospinal fluid MMP-9 levels.

BACKGROUND: Increasing evidence suggests seizures cause blood-brain barrier (BBB) dysfunction including decreased seizure threshold and higher onset potential of future seizures. However, the mechanisms underlying BBB damage in seizures remains poorly understood. Evidence in human and animal models shows BBB disruption is associated with activation of matrix metalloproteinase-9 (MMP-9) after cerebral ischemia and inflammation. The objective of this study was to determine whether MMP-9 concentrations in cerebral spinal fluid (CSF) are associated with BBB disruption in patients after epileptic seizures. METHODS: Thirty-one patients with generalized tonic-clonic (GTC) seizures were included in the study: 20 had recurrent GTC seizures (RS), and 11 had a single GTC seizure (SS) episode. Twenty-five adult non-seizure patients were used as controls. CSF samples were collected by lumbar puncture within 24 h after seizure cessation (range: 3-15 h, mean 6.2 h). CSF MMP-9 levels were determined by an enzyme-linked immunosorbent assay (ELISA). MMP enzyme activity was measured by gelatin zymography. The CSF/serum albumin ratio (albumin quotient, QAlb) was used as a measure of blood-brain barrier permeability. RESULTS: We found significantly higher CSF MMP-9 concentrations in seizure patients compared with controls (P < 0.001). CSF MMP-9 levels and QAlb values were higher in RS patients compared with SS and controls. Moreover, CSF MMP-9 concentration showed strong correlation between QAlb values (r = 0.76, P < 0.0001) and between CSF leukocyte counts (r = 0.77, P < 0.0001) in patients after seizures. Gelatin zymography showed MMP-9 proteolytic activity only in GTC seizure patients. CONCLUSIONS: Our results suggest MMP-9 plays a role in BBB dysfunction, characterized by invasion of leukocytes into the CSF during seizures.

Characteristics of physicochemical properties, structure and in vitro digestibility of seed starches from five loquat cultivars.

Starch plays a pivotal role in food and other industries, necessitating the exploration of new starch sources to cater the substantial requirement. This study delved into the variations in the physicochemical properties, structure attributes and in vitro digestibility of seed starches extracted from five distinct loquat cultivars (Eriobotrya japonica L.). The starch extraction yield of loquat seeds was found to be 45.2 % as an average. Loquat seed starches were designated as having high-amylose starch (>50 %). The starch granules exhibited similar shapes, but granular size significantly varied across the cultivars. Loquat starches presented a C-type crystalline pattern with relative crystallinity from 17.14 % to 21.06 %. The short-range ordered structure of the starches differed with loquat cultivars. The swelling power, solubility and amylose leaching of loquat starches increased with increasing temperature, significantly varying among different cultivars. Gelatinization parameters exhibited significant variations among the loquat starches. Different loquat starches exhibited pronounced variations in paste clarity, water and oil absorption capacity. Marked differences were detected in proportions of rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) across the five cultivars, with RS being particularly prominent fraction with an average of 84.30 %. These compressive findings offer valuable insights into the potential application of loquat seed starches in the formulation of foods and various industrial products.