Gut microbiota differences in stunted and normal-lenght children aged 36-45 months in East Nusa Tenggara, Indonesia.

The role of the gut microbiota in energy metabolism of the host has been established, both in overweight/obesity, as well as in undernutrition/stunting. Dysbiosis of the gut microbiota may predispose to stunting. The aim of this study was to compare the gut microbiota composition of stunted Indonesian children and non-stunted children between 36 and 45 months from two sites on the East Nusa Tenggara (ENT) islands. Fecal samples were collected from 100 stunted children and 100 non-stunted children in Kupang and North Kodi. The gut microbiota composition was determined by sequencing amplicons of the V3-V4 region of the 16S rRNA gene. Moreover, fecal SCFA concentrations were analyzed. The microbiota composition was correlated to anthropometric parameters and fecal metabolites. The phyla Bacteroidetes (Bacteroidota; q = 0.014) and Cyanobacteria (q = 0.049) were significantly higher in stunted children. Three taxa at genus levels were consistently significantly higher in stunted children at both sampling sites, namely Lachnoclostridium, Faecalibacterium and Veillonella (q < 7 * 10-4). These and 9 other taxa positively correlated to the z-score length-for-age (zlen), while 11 taxa negatively correlated with zlen. Several taxa also correlated with sanitary parameters, some of which were also significantly different between the two groups. All three fecal SCFA concentrations (acetate, propionate and butyrate) and their total were lower in stunted children compared to non-stunted children, although not significant for butyrate, indicating lower energy-extraction by the gut microbiota. Also, since SCFA have been shown to be involved in gut barrier function, barrier integrity may be affected in the stunted children. It remains to be seen if the three taxa are involved in stunting, or are changed due to e.g. differences in diet, hygiene status, or other factors. The observed differences in this study do not agree with our previous observations in children on Java, Indonesia. There are differences in infrastructure facilities such as clean water and sanitation on ENT and Java, which may contribute to the differences observed. The role of the gut microbiota in stunting therefore requires more in depth studies. Trial registration: the trial was registered at ClinicalTrials.gov with identifier number NCT05119218.

I am better than I look: genome based safety assessment of the probiotic Lactiplantibacillus plantarum IS-10506.

BACKGROUND: Safety of probiotic strains that are used in human and animal trials is a prerequisite. Genome based safety assessment of probiotics has gained popularity due its cost efficiency and speed, and even became a part of national regulation on foods containing probiotics in Indonesia. However, reliability of the safety assessment based only on a full genome sequence is not clear. Here, for the first time, we sequenced, assembled, and analysed the genome of the probiotic strain Lactiplantibacillus plantarum IS-10506, that was isolated from dadih, a traditional fermented buffalo milk. The strain has already been used as a probiotic for more than a decade, and in several clinical trials proven to be completely safe. METHODS: The genome of the probiotic strain L. plantarum IS-10506 was sequenced using Nanopore sequencing technology, assembled, annotated and screened for potential harmful (PH) and beneficial genomic features. The presence of the PH features was assessed from general annotation, as well as with the use of specialised tools. In addition, PH regions in the genome were compared to all other probiotic and non-probiotic L. plantarum strains available in the NCBI RefSeq database. RESULTS: For the first time, a high-quality complete genome of L. plantarum IS-10506 was obtained, and an extensive search for PH and a beneficial signature was performed. We discovered a number of PH features within the genome of L. plantarum IS-10506 based on the general annotation, including various antibiotic resistant genes (AMR); however, with a few exceptions, bioinformatics tools specifically developed for AMR detection did not confirm their presence. We further demonstrated the presence of the detected PH genes across multiple L. plantarum strains, including probiotics, and overall high genetic similarities between strains. CONCLUSION: The genome of L. plantarum IS-10506 is predicted to have several PH features. However, the strain has been utilized as a probiotic for over a decade in several clinical trials without any adverse effects, even in immunocompromised children with HIV infection and undernourished children. This implies the presence of PH feature signatures within the probiotic genome does not necessarily indicate their manifestation during administration. Importantly, specialized tools for the search of PH features were found more robust and should be preferred over manual searches in a general annotation.

Mono-Parasitic and Poly-Parasitic Intestinal Infections among Children Aged 36-45 Months in East Nusa Tenggara, Indonesia.

The prevalence of intestinal parasitic infection remains high in developing countries, especially because of geographic and socio-demographic factors. This study aimed to evaluate intestinal parasitic infection, as well as its risk factors, among children aged 36-45 months in a rural area (North Kodi) and an urban area (Kupang) of East Nusa Tenggara, Indonesia. Anthropometry, socio-demographic factors and personal hygiene practices were assessed. A total of 214 children participated in the study, and 200 stool samples were collected for intestinal parasite examination. Approximately 30.5% (61/200) of the children were infected with one or more intestinal parasites (67.2%; 41/61 being mono-parasitic infections and 32.8%; 20/61 being poly-parasitic infections). A total of 85 intestinal parasites were detected, consisting of 35.3% (30/85) protozoa and 64.7% (55/85) helminths. The predominant protozoa were Giardia lamblia (43%; 13/30) and Blastocystis spp. (33.3%; 10/30), whereas the predominant helminths were Trichuris trichiura (50.9%; 28/55) and Ascaris lumbricoides (43.6%; 24/55). Moreover, intestinal parasitic infection was associated with rural area (OR 4.5; 95%CI 2.3-8.6); the absence of treatment with deworming drugs (OR 2.56; 95%CI 1.3-5.0); sanitation facilities without a septic tank (OR 4.3; 95%CI 2.1-8.5); unclean water as a source of drinking water (OR 4.67; 95%CI 2.4-9.4); no handwashing practice after defecation (OR 3.2; 95%CI 1.4-7.3); and stunted children (OR 4.4; 95%CI 2.3-8.3). In conclusion, poly-parasitic infections were common in this study. Poor personal hygiene practice and sanitation factors contributed to the high prevalence of intestinal parasitic infection in 36-45-month-old children in East Nusa Tenggara, Indonesia.

The role of probiotics in the treatment of adult atopic dermatitis: a meta-analysis of randomized controlled trials.

BACKGROUND: Atopic dermatitis (AD) is chronic inflammatory skin disease that is relapsing and a serious condition that disrupts the quality of life of affected individuals. Probiotics are an immunomodulator that can enhance the immune control of atopic dermatitis. METHODS: All randomized controlled trials of probiotics for the treatment of adult AD published before December 2020 were included in this study from the PubMed databases and manual searching. RESULTS: Six randomized controlled trials (n = 241) were selected for this meta-analysis study. Probiotics were effective in treating adult patients with AD, indicated by the decrease in Scoring Atopic Dermatitis/SCORAD (Mean Difference (MD) - 7.90, 95% CI - 7.25 to - 6.92; p < 0.00001; I2 = 96%) and improved quality of life (MD - 7.68, 95% CI - 14.08 to - 1.29; p = 0.02; I2 = 47%) which were statistically significant. However, skin severity, itch severity, Dermatology Life Quality Index (DLQI), IL-4, TFN-γ, and IgE showed no significant difference in this meta-analysis study (p > 0.05). LIMITATIONS: The study found no available data for side effects of probiotics. STRENGTH: This meta-analysis analyzed a total of 241 AD patients of Asian and European origin. CONCLUSION: The use of probiotics decreased SCORAD significantly in adult patients with AD. Probiotics can improve the quality of life of patients with AD. The use of probiotics in atopic dermatitis has been widely studied, with controversial results. This meta-analysis suggests that the use of probiotics can improve SCORAD and the quality of life of patients with atopic dermatitis.

Effect of Different Functional Food Supplements on the Gut Microbiota of Prediabetic Indonesian Individuals during Weight Loss.

The gut microbiota has been shown in recent years to be involved in the development and severity of type 2 diabetes (T2D). The aim of the present study was to test the effect of a 2-week functional food intervention on the gut microbiota composition in prediabetic individuals. A randomized double-blind, cross-over trial was conducted on prediabetic subjects. Fifteen volunteers were provided products made of: (i) 50% taro flour + 50% wheat flour; (ii) these products and the probiotic L. plantarum IS-10506; or (iii) these products with beetroot adsorbed for a period of 2 weeks with 2 weeks wash-out in between. Stool and blood samples were taken at each baseline and after each of the interventions. The gut microbiota composition was evaluated by sequencing the V3-V4 region of the 16S rRNA gene and anthropometric measures were recorded. The total weight loss over the entire period ranged from 0.5 to 11 kg. The next-generation sequencing showed a highly personalized microbiota composition. In the principal coordinate analyses, the samples of each individual clustered closer together than the samples of each treatment. For six individuals, the samples clustered closely together, indicating a stable microbiota. For nine individuals, the microbiota was less resilient and, depending on the intervention, the beta-diversity transiently differed greatly only to return to the composition close to the baseline during the wash-out. The statistical analyses showed that 202 of the total 304 taxa were significantly different between the participants. Only Butyricimonas could be correlated with taro ingestion. The results of the study show that the highly variable interindividual variation observed in the gut microbiota of the participants clouded any gut microbiota modulation that might be present due to the functional food interventions.

Differences in immune status and fecal SCFA between Indonesian stunted children and children with normal nutritional status.

We recently showed that the gut microbiota composition of stunted children was different from that of children with normal nutritional status. Here, we compared immune status and fecal microbial metabolite concentrations between stunted and normal children, and we correlated macronutrient intake (including energy), metabolites and immune status to microbiota composition. The results show that macronutrient intake was lower in stunted children for all components, but after correction for multiple comparison significant only for energy and fat. Only TGF-ß was significantly different between stunted children and children of normal nutritional status after correction for multiple comparisons. TNF-alpha, IL-10, lipopolysaccharide binding protein in serum and secretory IgA in feces were not significantly different. Strikingly, all the individual short-chain and branched-chain fatty acids were higher in fecal samples of stunted children (significant for acetate, valerate and total SCFA). These metabolites correlated with a number of different microbial taxa, but due to extensive cross-feeding between microbes, did not show a specific pattern. However, the energy-loss due to higher excretion in stunted children of these metabolites, which can be used as substrate for the host, is striking. Several microbial taxa also correlated to the intake of macronutrients (including dietary fibre) and energy. Eisenbergiella positively correlated with all macronutrients, while an uncharacterized genus within the Succinivibrionaceae family negatively correlated with all macronutrients. These, and the other correlations observed, may provide indication on how to modulate the gut microbiota of stunted children such that their growth lag can be corrected. Trail registered at https://clinicaltrials.gov/ct2/show/NCT04698759.

Effect of Taro Starch, Beet Juice, Probiotic, and/or Psicose on Gut Microbiota in a Type 2 Diabetic Rat Model: A Pilot Study.

Background and Objectives. The gut microbiota has been shown to be involved in the development and severity of type 2 diabetes (T2D). The aim of the present study was to test the effect of potential functional food ingredients, alone or in combination, on the gut microbiota composition in diabetic rats in a pilot study of 1 week of feeding. Methods. In a pilot study to modulate the composition of the gut microbiota, (i) native taro starch, (ii) modified taro starch, (iii) beet juice, (iv) psicose, (v) the probiotic L. plantarum IS-10506, (vi) native starch combined with beet juice, (vii) native starch to which beet juice was adsorbed, (viii) modified starch combined with beet juice, and (ix) modified starch to which beet juice was adsorbed were fed to rats in which T2D was induced with streptozotocin (STZ). After one week, the composition of the gut microbiota was evaluated by sequencing the PCR-amplified V3-V4 region of the 16S rRNA gene. Results and Conclusions. The next-generation sequencing showed that 13 microbial taxa of the gut microbiota were significantly different between groups, depending on the treatment. The results of this pilot study will be used to design a 4-week intervention study in STZ-induced T2D rats to determine the best functional food for counteracting T2D, including their effects on satiety hormones. This should ultimately lead to the development of functional foods for prediabetic and diabetic individuals.

Gut microbiota profile of Indonesian stunted children and children with normal nutritional status.

The gut microbiota has been shown to play a role in energy metabolism of the host. Dysbiosis of the gut microbiota may predispose to obesity on the one hand, and stunting on the other. The aim of the study was to study the difference in gut microbiota composition of stunted Indonesian children and children of normal nutritional status between 3 and 5 years. Fecal samples and anthropometric measurements, in addition to economic and hygiene status were collected from 78 stunted children and 53 children with normal nutritional status in two regions in Banten and West Java provinces: Pandeglang and Sumedang, respectively. The gut microbiota composition was determined by sequencing amplicons of the V3-V4 region of the 16S rRNA gene. The composition was correlated to nutritional status and anthropometric parameters. Macronutrient intake was on average lower in stunted children, while energy-loss in the form of short-chain fatty acids (SCFA) and branched-chain fatty acids (BCFA) appeared to be higher in stunted children. In stunted children, at the phylum level the relative abundance of Bacteroidetes (44.4%) was significantly lower than in normal children (51.3%; p-value 2.55*10-4), while Firmicutes was significantly higher (45.7% vs. 39.8%; p-value 5.89*10-4). At the genus level, overall Prevotella 9 was the most abundant genus (average of 27%), and it was significantly lower in stunted children than in normal children (23.5% vs. 30.5%, respectively; q-value 0.059). Thirteen other genera were significantly different between stunted and normal children (q-value < 0.1), some of which were at low relative abundance and present in only a few children. Prevotella 9 positively correlated with height (in line with its higher relative abundance in normal children) and weight. In conclusion, Prevotella 9, which was the most abundant genus in the children, was significantly lower in stunted children. The abundance of Prevotella has been correlated with dietary fibre intake, which was lower in these stunted children. Since fibres are fermented by the gut microbiota into SCFA, and these SCFA are a source of energy for the host, increasing the proportion of Prevotella in stunted children may be of benefit. Whether this would prevent the occurrence of stunting or even has the potential to revert it, remains to be seen in follow up research.

Effect of functional food ingredients on gut microbiota in a rodent diabetes model.

BACKGROUND: The gut microbiota has been shown to be involved in the development and severity of type 2 diabetes. The aim of the present study was to test the effect of 4-week functional food ingredient feeding, alone or in combination, on the gut microbiota composition in diabetic rats. METHODS: Streptozotocin (STZ)-induced diabetic rats were treated for 4 weeks with (1) native taro starch, (2) modified taro-starch, (3) beet juice, (4) psicose, (5) the probiotic L. plantarum IS-10506, (6) native starch combined with beet juice, (7) native starch to which beet juice was adsorbed, (8) modified starch combined with beet juice or (9) modified starch to which beet juice was adsorbed, to modulate the composition of the gut microbiota. This composition was evaluated by sequencing the PCR amplified V3-V4 region of the 16S rRNA gene. RESULTS: The next-generation sequencing showed beneficial effects particularly of taro-starch feeding. Operational taxonomic units (OTUs) related to health (e.g. correlating with low BMI, OTUs producing butyrate) were increased in relative abundance, while OTUs generally correlated with disease (e.g. Proteobacteria) were decreased by feeding taro-starch. CONCLUSION: The results of study show that a 4-week intervention with functional food ingredients, particularly taro-derived starch, leads to a more healthy gut microbiota in rats that were induced to be diabetic by induction with STZ.

Modulation of Gut Microbiota Profile and Short-Chain Fatty Acids of Rats Fed with Taro Flour or Taro Starch.

To investigate the effect of flour and starch of the Indonesian native tuber "taro" on the composition and activity of the gut microbiota in diabetic rats, streptozotocin (STZ)-induced diabetic rats were fed normal chow (AIN), or AIN in which corn starch was replaced by either taro flour or purified taro starch for 4 weeks. Fecal samples were collected at baseline and after 4 weeks, and the composition of microbial communities was measured using 16S rRNA sequencing, while SCFAs were measured using ion chromatography. Bodyweight declined upon DM induction with STZ. Feeding taro starch led to a lower reduction in bodyweight than feeding taro starch, but this was only significant for taro starch in weeks 2, 3, and 4 (p = 0.02, p = 0.01, and p < 0.01, respectively). Both taro starch and taro flour induced changes in the gut microbiota composition compared to AIN, which were different for taro flour and taro starch. Bifidobacterium, Sutterella, and Prevotella were markers for taro flour feeding, while Anaerostipes was a marker for taro starch feeding. Induction of diabetes also led to changes in the microbiota composition. Random Forest correctly predicted for 16 of 18 samples whether rats were diabetic or not and correctly predicted 6 of 12 microbiota samples belonging to either taro flour- or taro starch-fed groups, indicating also some significant overlap in the substrate, as expected. Taro starch and taro flour both led to a significant increase in the fecal concentrations of acetate, propionate, and butyrate.

Low glycemic load after digestion of native starch from the indigenous tuber Belitung Taro (Xanthosoma sagittifolium) in a dynamic in vitro model of the upper GI tract (TIM-1).

BACKGROUND: Low glycemic foods are beneficial for people with type II diabetes. At the same time, sustained glucose release is also beneficial for people suffering from glycogen storage diseases. Taro (Xanthosoma sagittifolium) is a tuber indigenous to Indonesia, which has starch as the major storage carbohydrate. OBJECTIVE: The aim of the current study was to determine the speed of digestion of native and modified taro starch, compared to free glucose and wheat starch. DESIGN: This was investigated in a validated, dynamic computer-controlled in vitro model of the stomach and small intestine (TIM-1). Samples were taken from the dialysate, which reflected glucose absorbed in the blood stream. RESULTS: Native taro starch showed a ~1.5-fold reduced digestibility compared to glucose and a ~ 1.35-fold compared to wheat starch. In addition, digestion of native taro starch was moved towards the ileum, and later in time compared to glucose and wheat. With modified taro starch, these effects were not observed. CONCLUSION: In conclusion, native taro starch showed a lower glycemic load than wheat starch and modified taro starch and could be used as a substitute for refined foods by diabetics and people suffering from other glucose metabolic diseases.

Effect of the probiotic Lactobacillus plantarum IS-10506 on BDNF and 5HT stimulation: role of intestinal microbiota on the gut-brain axis.

BACKGROUND AND OBJECTIVES: Microbial communities residing in the gut play a major role in the communication between the gut and the brain through neural, immune, and hormonal routes. Changes in abundance of beneficial intestinal bacteria can affect health of individuals. Conversely, drugs, disease, diet and other factors can alter the gut microbiome. However, there is limited information on the effect of exogenous factors on gut microbiota. In this study, we investigated whether a beneficial bacterium, the probiotic Lactobacillus plantarum IS-10506, can stimulate the gut-brain axis using Wistar rats. MATERIALS AND METHODS: The animals were divided into two groups: one received L. plantarum IS strain 10506 supplementation, while the control group received no treatment. Activation of the gut-brain axis was evaluated by immunohistochemical analysis of intestinal and brain serotonin (5-HT) and brain neurotrophin (NT), serotonin transporter (5-HTT), and brain-derived neurotrophic factor (BDNF) levels. RESULTS: The results showed that BDNF (p< 0.000), NT (p< 0.000007), and 5-HTT (p< 0.000007) expression was upregulated in the brain along with intestinal 5-HT (p< 0.000) level in rats treated with L. plantarum strain IS-10506 relative to the control group. CONCLUSION: The probiotic L. plantarum IS-10506 stimulates the gut-brain axis and can potentially promote brain development and function.

Determinant of postprandial triglyceride levels in healthy young adults.

BACKGROUND: Fasting lipid profile does not necessarily illustrate the exact lipid dynamic in 24 h as human spends most of their time in postprandial state. Postprandial triglyceride (TG) has been reported to have advantages compared to fasting TG in terms of practicality and ability to predict cardiovascular events. This study aims to assess the determinant of postprandial TG in healthy young adults. METHODS: This is a quasy-experimental study that involved 200 healthy young adults. This study compared fasting with postprandial TG and analyzed the relationship between postprandial TG with various demographic and metabolic parameters after ingestion of standardized high fat liquid meal. RESULT: There was an upward trend from fasting TG to 2 h postprandial TG and 4 h postprandial TG. There was strong correlation between fasting TG and 2 h postprandial TG with 4 h postprandial TG (r = 0.731; p < 0.0001 dan r = 0.669; p < 0.0001, respectively). Whereas body mass index (BMI) and age showed weak correlation with 4 h postprandial TG (r = 0.141; p < 0.0001 dan r = 0.0747; p < 0.0001), fasting TG was the strongest predictor of 4 h postprandial TG (r = 0.669, B = 1.722 (95% CI 1.552 to 1.892), p < 0.0001). CONCLUSION: Fasting TG was the strongest determinant of 4 h postprandial TG in healthy young adults. We also observed strong correlation between 4 h postprandial TG and fasting TG. Hence, 4 h postprandial TG might potentially replaced fasting TG when measurement of fasting TG is not feasible.

Probiotic Lactobacillus plantarum IS 10506 supplementation increase SCFA of women with functional constipation.

BACKGROUND AND OBJECTIVES: Gut microbiota influences our health via multiple mechanisms. Microbiota produced Short Chain Fatty Acid (SCFA) as an energy to maintain gut ecosystem and physiology. Dysbiosis is correlated with SCFA imbalance which in turn resulted in physiological abnormalities in the intestine, such as functional constipation. MATERIALS AND METHODS: Randomized Double-Blind Controlled Trial (RCT) was conducted on women with functional constipation (n=37) in the community of Jakarta and profile of SCFA was assessed by using GC-MS from the stool after 21 days supplementation of fermented milk (placebo and probiotic). RESULTS: Probiotic supplementation significantly influenced acetate titer (p=0,032) marginally significant for propionate and butyrate (p=0.063 and p=0.068, respectively) and the respondent with increasing SCFA's metabolite are higher in probiotic group compared to the respondents in placebo group. Acetate is the highest SCFA titer found in faeces samples of women with functional constipation. CONCLUSION: Probiotic Lactobacillus plantarum IS 10506 supplementation influenced all the SCFA parameter (acetate, propionate and butyrate).

Lactobacillus plantarum IS-10506 probiotic administration increases amlodipine absorption in a rabbit model.

OBJECTIVE: Probiotics are beneficial in human health. In this study, we investigated the effect of probiotics on absorption of amlodipine, a dihydropyridine calcium antagonist used in the treatment of angina and hypertension, in a rabbit model. METHODS: Lactobacillus plantarum IS-10506 probiotic was administered for 14 days to male New Zealand rabbits. Blood samples were collected before and after probiotic supplementation. Amlodipine (10 mg) was then administered to all groups. Blood samples from a marginal vein were withdrawn at 5, 15, 30, 60, and 120 minutes to determine amlodipine concentrations in rabbit plasma. RESULTS: Amlodipine concentrations in the L. plantarum IS-10506 group were 4.95 ± 1.22, 8.71 ± 0.69, and 12.48 ± 2.53 ng/ml, and those in the control group were 1.69 ± 0.31, 3.89 ± 1.23, and 7.17 ± 1.85 ng/ml at 30, 60, and 120 minutes, respectively after administration of amlodipine. Amlodipine concentrations in the L. plantarum IS-10506 group were significantly higher than those in the control group at 30, 60, and 120 minutes after amlodipine administration. CONCLUSION: Our results suggested that supplementation of L. plantarum IS-10506 significantly increases amlodipine plasma concentrations in rabbits.

Development and validation of simple simultaneous analysis for amlodipine and glibenclamide by nonderivatization high-performance liquid chromatography-fluorescence.

Studies have shown that about 65% of diabetics have hypertension. Treatment for diabetic patients with hypertension is usually given a combination of drugs such as amlodipine (AML) and glibenclamide (GLI). The aim of this study was to develop and validate the simple simultaneous analysis method for separation of AML and GLI using high-performance liquid chromatography (HPLC) with fluorescence detector without derivatization. The arrangement of isocratic and gradient methods, mobile phase compositions, and flow rates to develop and validate the simple simultaneous analysis method for separation of AML and GLI by nonderivatization HPLC fluorescence was done. Optimum condition was obtained using an RP 18 (125 mm × 4 mm, i.d., 5 µm) and guard column RP 18 (4 mm × 4 mm, i.d., 5 µm) with mobile phase composition containing acetonitrile and phosphate buffer pH 3.0 using a 20:80 gradient condition at flow rate 1.0 ml/min measured at 361 nm for λ excitation and 442 nm for λ emission for AML and 235 nm for λ excitation and 354 nm for λ emission for GLI. The analysis of AML and GLI demonstrated a valid result with r 2 value 0.999, recoveries were 100.04% and 99.14% relative standard deviations were 0.508% and 0,797%, respectively, detection limits were 0.055 and 0.104 µg/ml, and quantification limits were 0.166 and 0.316 µg/ml, respectively. An accurate method of separation for AML and GLI using HPLC with fluorescence detector without derivatization has been validated.

The effect of an Annona muricata leaf extract on nutritional status and cytotoxicity in colorectal cancer: a randomized controlled trial.

BACKGROUND AND OBJECTIVES: Annona muricata leaf infusion has traditionally been consumed to maintain health, but is now considered for use in treating cancer patients. The objective of this study was to elucidate the effects of A. muricata leaf extract in humans and human cell lines. METHODS AND STUDY DESIGN: Thirty outpatients with colorectal cancer who had undergone primary tumor resection were enrolled in a randomized double-blind placebo- controlled pre-post-trial. They were divided into two groups: those who ingested A. muricata leaf extract (n=14) and those who ingested a placebo (n=14) daily for 8 weeks. Twenty-eight subjects completed the trial; they were equally distributed between the two groups. Serum from patients of both groups was compared for cytotoxicity against colorectal cancer cell lines. The nutritional status of patients was monitored throughout the study. RESULTS: Ex vivo and clinical studies showed higher cytotoxicity in the supplemented group compared with the placebo group. Further research is required to investigate the long-term effect of A.muricata leaf extract, particularly on parameters directly related to cytotoxic activity toward colorectal cancer cells and nutrition status.

From 'lactose intolerance' to 'lactose nutrition'.

The concept of lactose intolerance has become embedded in Western medicine and developing economy medicine. It is based on evidence that intestinal lactase activity persists into later childhood and throughout life in only a minority of the world's population, notably northern European-derived populations. These people have the T single nucleotide polymorphism (SNP) of the rs49882359 allele (C/T), also known as C/T-13910, the MCM6 gene which positively influences the lactase LCT gene. Other lactase persistent (LP) populations are found in Africa and the Middle East with different genetic variants. These SNPs represent co-evolution with dairying since the agricultural revolution and nutrient-dependent ecological adaptation. That said, gastrointestinal symptoms considered due to small intestinal lactose malabsorption are poorly correlated with lactase non-persistence (LNP), the situation for most people. With LNP, colonic microbiome lactase enables lactose fermentation to occur so that none is found in faeces. Whether the short chain fatty acids (SCFAs) and gases (hydrogen, carbon dioxide and methane) produced cause symptoms is dose-dependent. Up to 25 g of lactose at any one time can usually be consumed by a LNP person, but its food and meal pattern context, the microbiomic characteristics, age and other factors may alter tolerance. Thus, the notion that lactose intolerance is a disorder or disease of LNP people is misplaced and has been one of cultural perspective. What actually matters is whether a particular dairy product as normally consumed give rise to symptoms. It is, therefore, proposed that lactose tolerance tests be replaced with dairy food tolerance tests.

Traditional Indonesian dairy foods.

Indonesia is the largest archipelago blessed with one of the richest mega-biodiversities and also home to one of the most diverse cuisines and traditional fermented foods. There are 3 types of traditional dairy foods, namely the butter-like product minyak samin; yogurt-like product dadih; and cheese-like products dali or bagot in horbo, dangke, litsusu, and cologanti, which reflect the culture of dairy product consumption in Indonesia.

Effect of probiotic L. plantarum IS-10506 and zinc supplementation on humoral immune response and zinc status of Indonesian pre-school children.

A 90-day randomized, double-blind, placebo-controlled, pre-post trial was conducted in four groups of Indonesian children aged 12-24 months: placebo, probiotic, zinc, and a combination of probiotic and zinc (n=12 per group). Microencapsulated Lactobacillus plantarum IS-10506 of dadih origin was supplemented at a dose of 10(10)CFU/day as a probiotic. Zinc was supplemented as 20mg zinc sulfate monohydrate (8mg zinc elemental). Blood and stool samples were collected at baseline and at the end of the study period. Fecal sIgA was assessed by ELISA and serum zinc concentrations by ICP-MS. Fecal sIgA increased significantly in the probiotic group (30.33±3.32µg/g; p<0.01) and in the combination probiotic and zinc group (27.55±2.28µg/g; p<0.027), as compared with the placebo group (13.58±2.26µg/g). Changes in serum zinc concentrations in the combination probiotic and zinc group showed the highest elevation at the end of the study period. A combination of probiotic L. plantarum IS-10506 at a dose of 10(10)CFU/day and 8mg of elemental zinc supplementation showed a potential ability to improve the zinc status of pre-school children. Taken together, supplementation with the probiotic L. plantarum IS-10506 and zinc for 90 days resulted in a significantly increased humoral immune response, as well as improved zinc status, in young children.