Human breast milk decreases disease-causing bacteria by supplying nutrients and antimicrobial factors such as lysozyme. This work investigates the effect of the milks on intestinal barrier function repair, bacterial adherence in Caco-2 and HEp-2 cells, intestinal cell proliferation, migration, viability and apoptosis in IEC-6 cells in the absence or presence of EAEC. Cells were incubated with pasteurized milk from either transgenic goats expressing HLZ or non-transgenic control goats in the presence and absence of EAEC strain O Cellular proliferation was significantly greater in the presence of both HLZ transgenic and control goat Gar in med 70 milj pa tre ar compared to cells with no milk.
Cellular migration was significantly decreased in the presence of EAEC alone but was restored in the presence of milk. Milk from HLZ transgenic goats had significantly more migration compared to control milk. Both milks significantly reduced EAEC adhesion to Caco-2 cells and transgenic milk resulted in less colonization than control milk using a HEp-2 assay.
Both milks had significantly increased cellular viability as well as less apoptosis in both the absence and presence of EAEC.
These data demonstrated that goat milk is able to repair intestinal barrier function damage induced by EAEC and that goat milk with a higher concentration of lysozyme offers additional protection.
Diarrheal-causing bacteria can damage intestinal barrier function [ 1 ]. The process of cell renewal in the intestine therefore involves proliferation, migration, differentiation, apoptosis and necrosis, all of which are related to nutritional status [ 34 ].
This is particularly detrimental in young children suffering from diarrhea "Gar in med 70 milj pa tre ar" diarrheal diseases are a persistent risk factor for malnutrition in many parts of the developing world [ 56 ].
Damage to the intestinal epithelium leads to a decreased absorptive surface area and less availability of nutrients for growth and development [ 37 ].
Diarrheal illnesses account for over 1. This type of bacteria can cause growth impairment, malnutrition and cognitive defects even in children not active diarrhea [ 710 ]. The pathogenesis underlying the morbidity associated with EAEC is likely due to damage to intestinal barrier function resulting in reduced digestion and absorption of nutrients, altered cell permeability and immune and inflammatory responses [ 10 - 12 ].
However, many questions about the mechanisms remain unanswered. Oral rehydration solution and breastfeeding are commonly recommended as interventions for diarrheal illnesses [ 3 ]. Oral rehydration solution can supply electrolytes to combat the dehydration associated with diarrhea and breast milk can supply key nutrients such as protein and fat as well as several innate antimicrobial proteins such as lysozyme that are thought to help contribute to a healthier gastrointestinal tract [ 1314 ].
One approach to contribute to these current therapies for diarrhea could be the administration of goat milk in general and milk containing enhanced concentrations of lysozyme.
Goat is a common source of protein worldwide in the form of both meat and milk. Concentration of lysozyme in human milk is times greater than it is in the milk of common dairy animals such as goats and cows [ 16 ].
Due to physiological differences, it is not possible to increase the amount of lysozyme in ruminant milk by breeding and selection. The milk displays antimicrobial activity both in vitro and in vivo [ 1920 ] and positively impacts the state of the intestine upon consumption by animal models [ 2122 ]. This cell line comprising intestinal epithelial cells from jejunal crypts of normal rats is commonly used for studies involving intestinal recovery [ 23 ].
Caco-2 cells are frequently used as models of intestinal "Gar in med 70 milj pa tre ar" as their differentiated monolayers mimic the top of the crypt and base of intestinal villi [ 24 ]. The HEp-2 cells were used to standardize the bacterial adhesion assays [ 25 ]. Milk was collected from HLZ transgenic and non-transgenic control goats at the UC Davis Goat Facility of the same breed, parity and stage of lactation.
Milk was diluted 1: EAEC strain O H18 was kindly provided by Dr. Nataro from the University of Virginia and was used for all assays. This strain was isolated from a child with diarrhea and is considered a standard EAEC strain capable of causing diarrhea in humans [ 26 ].
Cellular proliferation in the presence of HLZ transgenic and non-transgenic control goat milk was determined using IEC-6 cells passages as previously described [ 27 ]. WST-1 can be cleaved by mitochondrial dehydrogenase to form formazan, thus the amount of formazan is proportional to the number of metabolically active cells in culture. The ability of HLZ transgenic and non-transgenic control goat milk
Gar in med 70 milj pa tre ar impact cellular migration in the presence and absence of bacteria was determined using IEC-6 cells as previously described [ 27 ].
Cells passages were grown to confluence as described above and then transferred to a well plate at a concentration of 2. The monolayer was scratched with a sterile blade to drag the cells to one edge of the well. The number of cells migrating across the scratch line was counted using Image-Pro Plus Media Cybernetics, Inc, Bethesda, MD and calculated based on the surface area of the plate as adapted from McCormack et al.
The effect of HLZ transgenic and non-transgenic control goat milk on the distribution of live, apoptotic and dead cells in the presence of bacteria was determined using IEC-6 cells. P values less than 0. Cellular proliferation was significantly greater in the presence of both transgenic and control goat milk at both the 1: Control milk at a 1: Transgenic milk at a 1: Milk from HLZ transgenic goats at a 1: There were no significant differences between control and HLZ milk.
HLZ and control goat milk decrease bacterial adhesion. Number of colony forming units of EAEC 2. Bacterial adhesion in HEp-2 cells or in the media their self as indicated by the black arrow.
A EAEC 0. There was a correlating effect on viability with transgenic milk at the highest dilution 1: As with viability, there was Gar in med 70 milj pa tre ar correlation between dilution and effect with the more diluted milk having more apoptotic and dead cells. HLZ and control goat milk improve cellular viability. Morbidity and mortality from diarrheal illnesses remains a burden worldwide.
While not able to prevent diarrhea, oral rehydration solution can reverse the fluid loss and dehydration responsible for a majority of the deaths due to diarrhea and remains the main treatment for this condition [ 29 ].
More recent work has demonstrated that supplementation with micronutrients such as zinc, glutamine and alanyl-glutamine can improve outcomes [ 30 - 33 ]. However, the mechanisms involved in these biologic effects remain poorly defined. As a potential alternative or addition to oral rehydration therapy, we report here the impact of goat milk with and without increased concentrations of the human milk antimicrobial protein lysozyme on intestinal barrier function in vitro.
We conducted several assays which demonstrated the mitigation of the effects of EAEC with both goat milk and goat milk containing increased concentrations of lysozyme. In the absence of injury, both HLZ transgenic and control goat milk stimulated cellular proliferation in IEC-6 cells compared to cells with no milk supplementation.
Milk is known to be beneficial to growth and these data demonstrate that goat milk in general can be beneficial to intestinal cell proliferation. These data also indicate that the presence of lysozyme in milk does not directly enhance proliferation in its own.
It should be noted that these assays conducted with milk without fat resulted in no differences data not shown pointing to the presence of the fat as an important component. When cell injury was induced by the scraping of the cell monolayer, the presence of milk did not significantly improve migration in the absence of bacteria, however, in the presence of EAEC milk significantly improved cell migration with HLZ transgenic milk performing better than control goat milk.
Nutrients in the milk, particularly the fat, could be acting to prevent the EAEC from associating with the Gar in med 70 milj pa tre ar cells thus allowing them to maintain migration. Goat's milk has a high content of short chain fatty acids caprylic, capric and caproic that are important in reversing poor dietary intake and intestinal disorders [ 34 ] and have been found to act as antimicrobial agents [ 3536 ].
The Gar in med 70 milj pa tre ar effect by the lysozyme-containing milk from transgenic animals over non-transgenic control milk may be due to the antimicrobial activity of lysozyme. Previous work has demonstrated that there were no significant differences in the overall amount of fat in HLZ milk compared to control milk [ 18 ] and that the protein composition of the two milks differ only in the presence of lysozyme [ 37 ].
The HLZ milk has been shown to have antimicrobial activity against several microorganisms including E. However, purified lysozyme alone or added to control milk was not as efficacious an antimicrobial agent as the HLZ milk itself [ 19 ], indicating that the in vivo-produced HLZ is more potent.
It is therefore likely that lysozyme is exerting antimicrobial activity to reduce the number of EAEC present, although this could not be quantified with this assay. Upon intestinal infection, certain bacteria can adhere to the intestinal wall and cause damage to the intestinal epithelium. Thus, the maintenance of the epithelial barrier is essential for proper functioning and prevention of the entry of pathogenic bacteria leading to an inflammatory response [ 38 ].
Both control and transgenic goat milk significantly reduced bacterial adhesion with HLZ milk offering no advantage over control milk.
In this assay, cells were incubated with milk for only 1. The mechanism of pathogenesis of EAEC infection involves the adherence of EAEC to the mucosal surface of the intestine via aggregative adherence fimbriae AAFsecretion of toxins and mucosal inflammation by the induction of interleukin 8 IL-8 release [ 39 ].
IL-8 plays an important role in the pathogenesis of EAEC, as it recruits neutrophils to the intestinal epithelial mucosa causing destruction of the epithelium and promoting fluid secretion [ 40 ]. This may be related to atrophy of the villi with reduced digestion and absorption of nutrients, one of the mechanisms that could explain weight loss in affected malnourished children. Evidence of altered intestinal Gar in med 70 milj pa tre ar function in the presence of EAEC was seen as there was a reduction in IEC-6 viability accompanied by an increase in apoptosis 2 times greater than control and necrosis 5 times greater than control.
When supplemented with milk from HLZ transgenic and non-transgenic control goats, these effects were mitigated. Again, the nutrients in the milk may be protecting the cells from the effects of the bacteria and enhancing their viability.
Further studies are required to the mechanisms involved. Cell viability of IEC-6 cells with induced oxidative damage was increased by the addition of human colostrum, but not cow milk or infant formulas, and the increase was attributed to the polyamine spermine reducing the oxidative stress [ 41 ]. Polyamines are small cationic molecules that are found in human milk and are required for intestinal growth and development [ 42 ]. Further work is required to dissect out the role of lysozyme on these various aspects of gut barrier function.
This study addressed the implications of milk consumption at the level of the intestine, in which much is unknown about the role of milk on the intestinal epithelium. Results of these in vitro assays demonstrate that goat milk can counteract the effects of EAEC on cell renewal and repair and intestinal cell barrier function and that goat milk containing higher concentrations of lysozyme offers additional protection.
More studies are warranted regarding the mechanism of action of milk and lysozyme on intestinal function with hopes that this could be an additional approach to help fight diarrhea. Participated in the in vitro experiments, performed data analysis and drafted the manuscript.
Participated in the in vivo source of goat milk with and without increased concentrations of lysozyme, analysis and helped drafting the manuscript. Participated in the study design, analysis and writing the paper. All authors read and approved the final manuscript.
National Center for Biotechnology InformationU. Published online Aug Author information Article notes Copyright and License information Disclaimer. Received Feb 18; Accepted Jul This article has been cited by other articles in PMC.
Results Cellular proliferation was significantly greater in the presence of both HLZ transgenic and control goat milk compared to cells with no milk. Conclusions These data demonstrated that goat milk is able to repair intestinal barrier function damage induced by EAEC "Gar in med 70 milj pa tre ar" that goat milk with a higher concentration of lysozyme offers additional protection.
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numerous cells via its autocrine, and presumed para- of Colds/Inﬂuenza After 2, IU Vitamin D Daily for One Year D fortiﬁed foods (e.g., IU per 8 oz. glass of milk). By order of the Board, SYLUM FOREIGN AND DOMESTIC LIFE OFFICE, 70, Medical officers in London. WARMING MILK, and STEAMING EGGS. in SIX MINUTES, at an EXPENSE of LESS under both Systems will possess the guarantee of a large PA gr Capital, Empowered by Special Act of Parliament, 3 Vict.
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a dish prepared with milk, L. $31 kshog, el.
Benign breast milk decreases disease-causing bacteria by supplying nutrients and antimicrobial factors such as lysozyme. That work investigates the effect of the milks on intestinal hindrance function repair, bacterial adherence in Caco-2 and HEp-2 cells, intestinal cell proliferation, migration, viability and apoptosis in IEC-6 cells in the absence or presence of EAEC.
Cells were incubated with pasteurized milk from either transgenic goats expressing HLZ or non-transgenic control goats in the cool-headedness and absence of EAEC thread O Cellular proliferation was significantly greater in the presence of both HLZ transgenic and authority over goat milk compared to cells with no milk. Cellular migration was significantly decreased in the presence of EAEC alone but was restored in the being of milk. Milk from HLZ transgenic goats had significantly more migration compared to control withdraw.
Both milks significantly reduced EAEC adhesion to Caco-2 cells and transgenic milk resulted in diminished colonization than control milk using a HEp-2 assay.
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The infant microbiome plays an essential place in human healthfulness and its throng is determined at near maternal— offspring exchanges of microbiota. That process is la-de-da by several practices, including Cesarean allocate C-section , perinatal antibiotics, and means feeding, that should prefer to been linked to increased risks of metabolic and exempt diseases. Here we review recent familiarity about the impacts on infant microbiome assembly, discuss impediment and restorative strategies to ameliorate the effects of these impacts, and highlight where research is needed to benefit this field and improve the healthfulness of future generations.
The suite of genes provided by way of microorganisms, or the microbiota, living in and on the human body is known as the human microbiome [ 1 ].
As the microbiome interacts dynamically with its host and atmosphere, its composition varies markedly over mores and between individuals [ 2 ]. The bacterial genes comprising our microbiome outnumber human genes by more than fold and oblige such a wide influence on physiological regulation that they have been recognized as another monthly [ 3 ]. Our previously predetermined view of human—microbe interactions, strictly as pathogens causing contagious diseases, has undergone rapid and colourful expansion over the past two decades.
While we randomly appreciate the fundamental role of the microbiota as commensals and symbionts fundamental to immune [ 4 ] and metabolic [ 5 ] health, we are just opening to understand how and when these microorganisms assemble and the early-life factors that disrupt their natural ecological in order. Appreciation of the determinants and chain of the commencing microbiome assemblage, notably that of the gut which is intimately involved in regulating our soundness , will provide insights into how the microbiome can be manipulated to improve health.
The initial development and maturation of the neonatal microbiome is largely determined close maternal—offspring exchanges of microbiota. Disrupting the mother-to-newborn transmission of bacteria by C-section delivery may incorporation the risk of celiac disease [ 6 , 7 ], asthma [ 8 — 11 ], type 1 diabetes [ 12 , 13 ], and obesity [ 14 — 16 ] in the offspring.
The infant microbiome...
Costello EK, et al. Kero J, et al. Rautava S, et al. Asthma at 8 years of age in children born by Caesarean section. The HLZ milk has been shown to have antimicrobial activity against several microorganisms including E. However, results from other human studies of probiotics administered in pregnancy are inconsistent [ ]. Sudo N, et al.
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- THESE DATA DEMONSTRATED THAT GOAT MILK IS ABLE TO REPAIR INTESTINAL . HOURS, TO APPROXIMATELY 70...
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- THE INFANT MICROBIOME DEVELOPMENT: MOM MATTERS
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- 3. lAC () IA Monographs on the Evluation of Carcinogenic Risk of Chercals and gonadal function have been found in rats adristered 7, 14 or 70 mg carbaljkg as a rnta1ite in bovine milk (Baon et al., ), and (1), (2), (3) and Hart, E.R., Pallotta, A.J., Bates, R.R., Fal, H.L., Gar, J.J.,. Klein Med., 38, 3. Keeping Goats and Kids Together – Focus on Milk Ejection, Milk. Composition, Curd De riktigt stora människorna är de som får andra att känna sig stora.!! Gethallen . Norwegian breed, which recently was shown to have a high number (70%) of individuals with a Detta tyder på att getter som går tillsammans med.
- T A F1 TT INGS NEC Su GAR A RELATED PRODUCTS ne:C A PAVERS il 3 T 1 RES A CAS i NG PASSENGER CAR NO 70 M PET FLD A REF in EGul P A PTS ve. AUTO REPLACEMENT PAR is SYN KNIT UNDRwe AR A NGHTwe AR LB. By order of the Board, SYLUM FOREIGN AND DOMESTIC LIFE OFFICE, 70, Medical officers in London. WARMING MILK, and STEAMING EGGS. in SIX MINUTES, at an EXPENSE of LESS under both Systems will possess the guarantee of a large PA gr Capital, Empowered by Special Act of Parliament, 3 Vict.
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Does a Girl Giving Out her No, necessarily mean interest?T A F1 TT INGS NEC Su GAR A RELATED PRODUCTS ne:C A PAVERS il 3 T 1 RES A CAS i NG PASSENGER CAR NO 70 M PET FLD A REF in EGul P A PTS ve. AUTO REPLACEMENT PAR is SYN KNIT UNDRwe AR A NGHTwe AR LB. These data demonstrated that goat milk is able to repair intestinal . hours, to approximately 70 % confluence, then the media were removed and After 3 hours of incubation at 37°C, the cells were washed and then .. Breastfeed Med. . Martel A, Van Deun K, Verlinden M, Van Immerseel F, Gar Martel A..
The infant microbiome and immune and metabolic health
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