UV-vis absorbance spectra, molar extinction coefficients and circular dichroism spectra for the two cyanobacterial metabolites anabaenopeptin A and anabaenopeptin B.

The UV-vis absorbance spectra, molar extinction coefficients and circular dichroism spectra, as well as NMR and high resolution tandem mass spectrometry spectra were determined for two prominent secondary metabolites from cyanobacteria, namely anabaenopeptin A and anabaenopeptin B. The compounds were extracted from the cyanobacterium CBT929 and purified by flash chromatography and HPLC. Exact amounts of isolated compounds were assessed by quantitative H-NMR with internal calibrant ethyl 4-(dimethylamino)benzoate in DMSO‑ at 298 K with a recycle delay (d1) of 120 s.

Shifts in microbial community structure of granular and liquid biomass in response to changes to infeed and digester design in anaerobic digesters receiving food-processing wastes.

There have been few studies, to date, examining the effect of seed sludge on the microbial community established in a new anaerobic digestion (AD) system and whether or not the population present in the seed sludge establishes it self as the predominant population. Further, no reported studies have yet examined the differences in microbial populations that result from the formation of granular biomass in upflow anaerobic sludge blanket (UASB) systems. This study focused on examining the changes in microbial diversity between the initial seed sludge and the community that becomes established in a new digester.

Inflammatory responses in epithelia: endotoxin-induced IL-6 secretion and iNOS/NO production are differentially regulated in mouse mammary epithelial cells.

Background: IL-6 is a pro-inflammatory cytokine that signals via binding to a soluble or membrane bound receptor, while nitric oxide (NO), an oxidative stress molecule, diffuses through the cell membrane without a receptor. Both mediators signal through different mechanisms, yet they are dependent on NFκB. We proposed that both mediators are co-induced and co-regulated in inflamed mammary epithelial cells.

Putting microbes to work in sequence: recent advances in temperature-phased anaerobic digestion processes.

Methane biogas production through anaerobic digestion (or biomethanation) is one of the few technologies that both produce bioenergy and protect the environment. When the focus of anaerobic digestion (AD) is shifted from primarily wastewater treatment to bioenergy production, efficiency and process stability become critical to the economic viability of AD technologies. Temperature-phased anaerobic digestion (TPAD) is a promising process that can significantly enhance both digestion efficiency and process robustness.

Evaluations of different hypervariable regions of archaeal 16S rRNA genes in profiling of methanogens by Archaea-specific PCR and denaturing gradient gel electrophoresis.

Different hypervariable (V) regions of the archaeal 16S rRNA gene (rrs) were compared systematically to establish a preferred V region(s) for use in Archaea-specific PCR-denaturing gradient gel electrophoresis (DGGE). The PCR products of the V3 region produced the most informative DGGE profiles and permitted identification of common methanogens from rumen samples from sheep. This study also showed that different methanogens might be detected when different V regions are targeted by PCR-DGGE.

Lactoferrin interaction with retinoid signaling: cell growth and apoptosis in mammary cells.

Lactoferrin (Lf) is a multifunctional iron-binding protein that was first identified in mammary secretions, but is synthesized by most mammalian tissues. The protein has a signal sequence that dictates secretion; it also has a nuclear localization sequence that facilitates entry into the cell nucleus. The mechanism of the latter action is currently unknown, but is thought to occur via a Lf receptor.

In vivo effects of bifidobacteria and lactoferrin on gut endotoxin concentration and mucosal immunity in Balb/c mice.

The aim of the present study was to examine the effects of oral supplementation of newborn Balb/c mice with bifidobacteria (B. infantis, B. bifidum) and iron-free apo-lactoferrin (bovine, human) on gut endotoxin concentration and mucosal immunity.

In vitro growth responses of bifidobacteria and enteropathogens to bovine and human lactoferrin.

A series of in vitro experiments was performed to test the ability of bovine and human lactoferrin to influence the growth of the gram-positive probiotic bacteria, Bifidobacterium bifidum, Bifidobacterium infantis, and Lactobacillus acidophilus, as well as the gram-negative enteric bacteria, E. coli O157:H7 and Salmonella typhimurium. None of the lactoferrin preparations stimulated the growth of the tested strains.

Bovine lactoferrin binds to insulin-like growth factor-binding protein-3.

Insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) has been shown to have IGF independent actions that appear to be mediated by specific IGFBP-3 binding proteins located on cell membranes. We show here using Western ligand blotting, a number of mammary membrane proteins that bind 125I-labeled rhIGFBP-3. Immunoprecipitation studies demonstrated that the >70 kDa protein was identified from bovine mammary microsomes as bovine lactoferrin (bLf).

Introduction of the human growth hormone gene into the guinea pig mammary gland by in vivo transfection promotes sustained expression of human growth hormone in the milk throughout lactation.

We tested the feasibility of transfecting mammary tissue in vivo with an expression plasmid encoding the human growth hormone (hGH) gene, under the control of the cytomegalovirus promoter. Guinea pig mammary glands were transfected with plasmid DNA infused through the nipple canal and expression was monitored in control and transfected glands by radioimmunoassay of milk samples for hGH. Sustained expression of hGH throughout lactation was attained with a polyion transfection complex shown to be optimal for the transfection of bovine mammary cells, in vitro.

Effect of substratum on growth, cell morphology and lactoferrin synthesis and secretion in bovine mammary cell culture.

The role of extracellular matrix in morphology, growth and lactoferrin synthesis and secretion in bovine mammary cells from a developing gland is poorly defined. In this study, bovine mammary cells from a hormone-primed developing gland were isolated and cultured on plastic, collagen, embedded within collagen, or on EHS-matrix, with the hormones prolactin, insulin, and cortisol in the presence or absence of fetal calf serum. Mammary cells on plastic or collagen spread and formed confluent cells sheets, while those embedded within collagen or on EHS-matrix maintained their acinar-like structure.

Cloning of bovine parathyroid hormone-related protein (PTHrP) cDNA and expression of PTHrP mRNA in the bovine mammary gland.

Parathyroid hormone-related protein (PTHrP) produced by the mammary gland has been postulated to have multiple functions in both the mother and neonate. In humans, alternative 3'-mRNA splicing and endoproteolytic processing result in multiple bioactive PTHrP peptides. Multiple PTHrP peptides also have been reported in bovine milk.

In vitro model of parathyroid hormone-related protein secretion from mammary cells isolated from lactating cows.

Parathyroid hormone-related protein (PTHrP) is produced by the lactating mammary gland and is present in milk in a biologically active form. The goal of this investigation was to determine if cells cultured from the lactating mammary glands of cows would secrete PTHrP in vitro. Mammary acini were isolated from lactating cows at 1-6 wk after calving, and fresh or cryopreserved mammary acini were cultured for 14 d on Type I collagen.

Structure and biological actions of lactoferrin.

Lactoferrin is an iron-binding glycoprotein of the transferrin family, first isolated from milk but also found in most exocrine secretions as well as in the secondary granules of neutrophils. The many reports on its antimicrobial and antiinflammatory activity in vitro identify lactoferrin as important in host defense against infection and excessive inflammation. Most if not all lactoferrin actions are mediated through iron sequestration and/or interaction with a large variety of ligands including microbial cell wall components and cellular receptors, through its highly positively charged N-terminus.

Developmental regulation and partial characterization of growth factors in the bovine mammary gland.

Bovine mammary gland secretions from several developmental stages were shown to stimulate [3H]thymidine incorporation into AKR-2B mouse embryo fibroblast cells. In virgin heifers, mammary secretions at 1% concentration stimulated thymidine incorporation into AKR-2B cells more than threefold compared with 10% (v/v) fetal calf serum. The growth-promoting activity peaked at an early stage of the last trimester (7-8 months) of gestation and declined after this until the colostrum forming stage.

Morphological and functional differentiation of cryopreserved lactating bovine mammary cells cultured on floating collagen gels.

Cryopreserved bovine mammary epithelial cells prepared from lactating mammary tissue synthesize and secrete the milk proteins alpha s1-casein, lactoferrin (Lf), and alpha-lactalbumin during in vitro culture on collagen gels in serum-free medium. Each milk protein is differently regulated by detachment and thickness of the collagen substratum, fetal calf serum, and prolactin in the medium. Collagen detachment did not modulate lactoferrin secretion but strongly induced casein secretion, with detachment on day 6 (after formation of cell sheets) inducing casein secretion to 3 micrograms/ml medium, which was 2-3-fold higher than for cells on collagen detached on day 2 (prior to cell spreading to form sheets), and ten-fold higher than for cells grown on collagen not detached.

Bovine mammary lactoferrin: implications from messenger ribonucleic acid (mRNA) sequence and regulation contrary to other milk proteins.

The regulation of bovine mammary lactoferrin, an important component of the antimicrobial defenses of the mammary gland, is poorly understood compared with the other milk proteins. The complete sequence for bovine lactoferrin mRNA shows it to be highly homologous to other lactoferrins and transferrins. However, regional differences in the deduced AA sequence of bovine lactoferrin compared with human lactoferrin and transferrin imply functional differences between them.

Bovine lactoferrin mRNA: sequence, analysis, and expression in the mammary gland.

The mRNA sequence for bovine lactoferrin expressed in the mammary gland was determined by sequencing three over lapping cDNA clones and by direct sequencing of the mRNA. The mRNA (2351 bases) codes for a 708 amino acid protein with a 19 amino acid signal peptide immediately preceding a sequence identical to the N-terminal 40 amino acids reported for bovine lactoferrin. A putative destabilizing sequence (AUUUA) was identified in the 3'-untranslated region.

Dietary fat composition influences fatty acid composition of milk fat globule membrane in lactating cows.

Milk fat globule membranes are derived directly from the apical plasma membrane of mammary epithelial cells. To evaluate the effect of dietary fat on mammary membranes, we determined the fatty acid composition of the milk fat globule membrane in lactating dairy cows fed diets supplemented with fats of different fatty acid composition, or infused intravenously with soy oil emulsion. A preliminary survey, using an abbreviated preparation procedure (membranes isolated at 48,000 x g-max for 15 min), yielded about 45% of the total membrane fatty acids that could be recovered by centrifuging at the same speed for 120 min, and showed that changes in fatty acid composition of membranes reflected dietary fatty acids to some extent.

In vitro culture of cryopreserved bovine mammary cells on collagen gels: synthesis and secretion of casein and lactoferrin.

The preparation, cryopreservation, and culture on type I collagen gels of lactating bovine mammary cells with prolonged milk protein synthesis and secretion in vitro is described. Cryopreserved cells prepared as acinar fragments from either lactating or developing mammary glands attached to the collagen substratum within 24-48 hr after plating in serum and hormone supplemented medium. During continued culture in hormone-supplemented (insulin, cortisol, and prolactin) serum-free medium outgrowth of cells from the attached acinar fragments was observed beginning on day 2, with continued outgrowth to near confluence by day 6.

Effect of dietary fat and lactation status on insulin binding to bovine milk fat globule membranes.

Lactating cows were used to examine the relationship between lactation status and insulin binding to milk fat globule membranes. Variables evaluated were daily milk yield, stage of lactation, breed, age, lactation number, daily milk fat and protein yields, milk fat and protein percentages, breeding status, body weight, body weight.75, and mammary health.

Serum isocitrate dehydrogenase during polybrominated biphenyl toxicosis in dairy cattle.

Bovine serum isocitrate dehydrogenase (sICDH) was investigated in dairy cattle as a clinical measurement indicative of hepatic injury. Conditions for optimization of isocitrate dehydrogenase assays for bovine serum are described. Assays of sICDH in normal cattle show average activities of .

Characterization of insulin binding to bovine liver and mammary microsomes.

Bovine liver and mammary gland (MG) appear metabolically independent of insulin, yet the specificity and kinetics of 125I-insulin (125I-INS) binding to bovine liver and MG microsomes (MIC) indicate the presence of insulin receptors in MIC from both tissues. The insulin receptors from bovine liver (Kd = 7.6 X 10(-10) M) and MG (Kd = 9.

Effects of polybrominated biphenyls on the excretion of steroids.

When polybrominated biphenyls (fireMaster BP-6, PBB) are ingested by cattle, they have been shown to alter hepatic enzyme systems, and produce renal lesions with chronic high exposure. These changes provide mechanisms for alteration of the metabolism and clearance of steroid hormones that might then affect reproductive function. This study was conducted to examine the effects of PBB on the excretion of radiolabel from injected estradiol-17 beta and progesterone.