The human embryonic genome is karyotypically complex, with chromosomally abnormal cells preferentially located away from the developing fetus.

STUDY QUESTION: Are chromosome abnormalities detected at Day 3 post-fertilization predominantly retained in structures of the blastocyst other than the inner cell mass (ICM), where chromosomally normal cells are preferentially retained? SUMMARY ANSWER: In human embryos, aneuploid cells are sequestered away from the ICM, partly to the trophectoderm (TE) but more significantly to the blastocoel fluid within the blastocoel cavity (Bc) and to peripheral cells (PCs) surrounding the blastocyst during Day 3 to Day 5 progression. WHAT IS KNOWN ALREADY: A commonly held dogma in all diploid eukaryotes is that two gametes, each with 'n' chromosomes (23 in humans), fuse to form a '2n' zygote (46 in humans); a state that remains in perpetuity for all somatic cell divisions. Human embryos, however, display high levels of chromosomal aneuploidy in early stages that reportedly declines from Day 3 (cleavage stage) to Day 5 (blastocyst) post-fertilization. While this observation may be partly because of aneuploid embryonic arrest before blastulation, it could also be due to embryo 'normalization' to a euploid state during blastulation. If and how this normalization occurs requires further investigation. STUDY DESIGN, SIZE, DURATION: A total of 964 cleavage-stage (Day 3) embryos underwent single-cell biopsy and diagnosis for chromosome constitution. All were maintained in culture, assessing blastulation rate, both for those assessed euploid and aneuploid. Pregnancy rate was assessed for those determined euploid, blastulated and subsequently transferred. For those determined aneuploid and blastulated (174 embryos), ICM (all 174 embryos), TE (all 174), Bc (47 embryos) and PC (38 embryos) were analyzed for chromosome constitution. Specifically, concordance with the original Day 3 diagnosis and determination if any 'normalized' to euploid karyotypes within all four structures was assessed. PARTICIPANTS/MATERIALS, SETTING, METHODS: All patients (144 couples) were undergoing routine preimplantation genetic testing for aneuploidy in three IVF clinical settings. Cleavage-stage biopsy preceded chromosome analysis by next-generation sequencing. All patients provided informed consent. Additional molecular testing was carried out on blastocyst embryos and was analyzed for up to four embryonic structures (ICM, TE, Bc and PC). MAIN RESULTS AND THE ROLE OF CHANCE: Of 463/964 embryos (48%) diagnosed as euploid at Day 3, 70% blastulated (leading to a 59% pregnancy rate) and 30% degenerated. Conversely, of the 501 (52%) diagnosed as aneuploid, 65% degenerated and 35% (174) blastulated, a highly significant difference (P < 0.0001). Of the 174 that blastulated, the ratio of '(semi)concordant-aneuploid' versus 'normalized-euploid' versus 'other-aneuploid' embryos was, respectively, 39%/57%/3% in the ICM; 49%/48%/3% in the TE; 78%/21%/0% in the PC; and 83%/10%/5% in the Bc. The TE karyotype therefore has a positive predictive value of 86.7% in determining that of the ICM, albeit with marginally higher aneuploid rates of abnormalities (P = .071). Levels of abnormality in Bc/PC were significantly higher (P < 0.0001) versus the ploidy of the ICM and TE and nearly all chromosome abnormalities were (at least partially) concordant with Day 3 diagnoses. LIMITATIONS, REASONS FOR CAUTION: The results only pertain to human IVF embryos so extrapolation to the in vivo situation and to other species is not certain. We acknowledge (rather than lineage-specific survival, as we suggest here) the possibility of other mechanisms, such as lineage-specific movement of cells, during blastulation. Ethical considerations, however, make investigating this mechanism difficult on human embryos. WIDER IMPLICATIONS OF THE FINDINGS: Mosaic human cleavage-stage embryos can differentiate into a euploid ICM where euploid cell populations predominate. Sequestering of aneuploid cells/nuclei to structures no longer involved in fetal development has important implications for preimplantation and prenatal genetic testing. These results also challenge previous fundamental understandings of mitotic fidelity in early human development and indicate a complex and fluid nature of the human embryonic genome. STUDY FUNDING/COMPETING INTEREST(S): This research was funded by Organon Pharmaceuticals and Merck Serono by grants to W.G.K. W.G.K. is also an employee of AdvaGenix, who could, potentially, indirectly benefit financially from publication of this manuscript. R.C.M. is supported by the National Institute of General Medical Sciences of the National Institutes of Health under award number R35GM133747. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. D.K.G. provides paid consultancy services for Care Fertility. TRIAL REGISTRATION NUMBER: : N/A.

Susceptibility of Campylobacter jejuni to organic acids and monoacylglycerols.

Organic acids can be used as feed supplements or for treatment of poultry carcasses in processing plants. The antimicrobial activity of nineteen organic acids and two monoacylglycerols in cultures of Campylobacter jejuni CCM 6214(T) (ATCC 33560) was determined using a SYBR Green-based real-time PCR assay. The IC(50) was a concentration at which only 50 % of a bacteria specific DNA sequence was amplified. Caprylic, capric and lauric acids were the most efficient antimicrobials among the compounds tested (IC(50) < or = 0.1 mg/mL). In a weakly acidic environment (pH 5.5), the antimicrobial activity was more pronounced than at pH 6.5. At pH 5.5, oleic and fumaric acid also had clear antimicrobial activity, as did monocaprylin. The antimicrobial activity of acetic, butyric, stearic and succinic acid was low. In cells treated with fumaric acid, the potential of potassium and tetraphenylphosphonium ion-selective electrodes changed, indicating an increase in cytoplasmic and outer membrane permeability, respectively. No changes in membrane permeability were observed in cells treated with capric acid or monocaprin. Transmission electron microscopy revealed separation of the inner and outer membrane in cells treated with capric and fumaric acid, as well as cytoplasmic disorganization in cells exposed to capric acid.

Gestational diabetes insipidus, HELLP syndrome and eclampsia in a twin pregnancy: a case report.

We report a case of eclampsia in a twin pregnancy complicated by HELLP syndrome and diabetes insipidus. This confluence of disease processes suggests that a modification of common magnesium sulfate treatment protocols may be appropriate in a certain subset of patients.

Serous cystadenoma with omental caking and ovarian torsion: an unusual case presentation.

BACKGROUND: This case evaluates a patient with abdominal pain who presented with a pelvic mass and imaging studies suspicious for malignancy. CASE: A 21-year-old, gravida 0, para 0, was admitted after presenting to the outpatient with abdominal pain. Ultrasound revealed a large 17 x 20 cm pelvic mass with solid and cystic components. CT scan revealed worrisome findings including the finding of "omental caking" and nodal enlargement. CA 125 was elevated at 85 U/ml. Final pathology after surgical removal of the mass showed evidence of serous cystadenoma with ovarian torsion without signs of malignancy. The patient had an unremarkable postoperative course and was discharged in stable condition. CONCLUSION: Although uncommon, pelvic masses that are benign may mimic malignant masses with extradnexal inflammation.

Fermentation of carbohydrates and yield of microbial protein in mixed cultures of rabbit caecal microorganisms.

Fermentation pattern and yields of microbial protein were investigated in cultures of the rabbit caecal contents supplied with glucose, xylose, starch, pectin and xylan. Rabbits at the age of 4 weeks (before weaning) and 3 months were slaughtered, their caecal contents added at 1.1% to growth media and incubated anaerobically at 39 degrees C for 18 h. Caecal microorganisms of 4-week-old rabbits produced no methane and caproate, less butyrate, but more propionate than microorganisms of 3-month-old rabbits. In both groups of rabbits, fermentation of xylose produced significantly more propionate and less butyrate than fermentation of glucose. More propionate and less acetate was formed from starch than from pectin. In caecal cultures from 4-week-old rabbits with pectin, the molar percentages of acetate was significantly higher and percentages of other short-chain fatty acids (SCFA) lower than in cultures with starch or xylan. In cultures from 3-month-old rabbits, fermentation of pectin and xylan produced similar SCFA profiles, different from SCFA molar composition in cultures with starch. Average production of microbial protein was 129 mg per 1 g of carbohydrate digested (range 110 to 141 mg/g). Protein yields were the same on glucose and xylose, but nonsignificantly higher on starch than on pectin and xylan. It can be concluded that the characteristics of substrate affected fermentation pattern in mixed cultures of rabbit caecal microorganisms. Substrate effects on protein yields were not statistically significant, due to high variation.

Heat-induced formation of soluble Maillard reaction products and its influence on utilization of glucose by rumen bacteria.

In a series of experiments with pure strains of rumen bacteria the effect of heat-induced formation of soluble Maillard reaction products on the utilization of glucose was examined. Maillard reaction products were prepared from glucose and amino acids, which were dissolved in a growth medium and autoclaved at 100 degrees C and 120 degrees C. Glucose was utilized almost completely in all cultures, no matter whether it was bound in Maillard products or not. The complexing of glucose with amino acids lowered the growth rate and growth yields in most rumen bacteria studied. In some strains, however, the effect of the Maillard reaction was small. It was concluded that the heat damage of carbohydrates in feedstuffs could be attributed to final stages of the Maillard reaction (formation of insoluble polymers), rather than to initial ones.

Influence of redox potential on metabolism of glucose in mixed cultures of rumen microorganisms.

The rumen inocula taken from cows and sheep were incubated in pH controlled anaerobic batch cultures and the effect of the elevated redox potential (Eh) on metabolism of glucose was examined. In Eh regulated cultures (Eh = +100 mV) the VFA production and production of methane were decreased with a concomitant increase of production of lactate. In these cultures counts of lactilytic bacteria tended to be lower and counts of lactic acid bacteria higher than in cultures with a normal Eh. The elevated Eh also exerted an inhibitory effect on the rumen protozoa. The relevance of these changes to the rumen metabolism is discussed.

[Determination of aflatoxin M1 level in milk in the production of baby and children's food using immunoassay]. / Verfolgung des Aflatoxin-M1-Gehaltes in Milch für die Produktion von Süglings- und Kindernahrung mittels Immunoassay.

Using commercial immunokits the concentration of aflatoxin M1 was measured in 376 samples of raw milk from farms in the area of a new dairy plant producing milk baby foods. 87.8% of the samples contained no aflatoxin M1 (detection limit 0.025 micrograms/l) and only 2 samples (0.5%) possessed higher concentration than 0.1 microgram/l, which represents the tolerance limit for aflatoxin M1 in baby milk foods admitted in Czechoslovakia.

[Immunobiologic properties of lactobacilli]. / Imunobiologické vlastnosti laktobacilov.

The authors investigated in experiments on mice immunobiological properties of selected strains of lactobacilli (Lbc. acidophilus, Lbc. casei and Lbc. delbruecki). Their immunostimulating action was evaluated from the migrating capacity of lymphocytes into the interepithelial spaces and lamina propria mucosae of the gut. The most marked changes were observed in the group of animals to whom Lbc. acidophilus and Lbc. casei was administered for two weeks by a gastric tube. The protective properties of lactobacilli on the course and development of model infections (virus of encephalomyocarditis) was greatest in mice given Lbc. casei and Lbc. acidophilus by the intraperitoneal route four days before infection. At the end of the two-week period in the Lbc. casei group 66% mice survived and in the Lbc. acidophilus group 34%. The ability of lactobacilli to influence the interferon producing activity was investigated in vitro on a model of peritoneal cells obtained from premedicated mice. The lactobacilli strains themselves did not have interferon inducing properties. However, when the interferon producing capacity of peritoneal cells was assessed after administration of the viral inducer (virus of Newcastle disease) the capacity was much higher, when compared with controls.

Interferences in radioimmunoassay of aflatoxins in food and fodder samples of plant origin.

Cross-reactions and resulting nonspecific binding of substances with structures resembling aflatoxins (derivatives of coumarin, and cinnamonic and benzoic acids, etc.) were investigated. The concentrations of these substances causing erroneously high or false positive values in radioimmunoassay were determined. One microgram aflatoxin B1/kg sample may be simulated by the occurrence of 5 g coumarin, 10 g caffeic acid, 16 g chlorogenic acid, or 15 g vanillin/kg fodder or food sample.

Production of acids from inulin by a mixed culture of rumen microorganisms.

The production of volatile fatty acids (VFA) and lactic acid from inulin (plant fructan with 2-1 bonds) in in vitro rumen fermentations was followed. The experiments were performed with inocula from wethers receiving two rations, with two pH regimes and using as inocula either whole rumen contents or the corresponding rumen fluid. The following results were obtained: The VFA production was higher when using inocula from hay and concentrate fed wethers. In contrast to this, the lactic acid production was higher when using inocula from hay fed wethers. The VFA production tends to be higher in a weakly acid medium than in a neutral medium. Acetate-to-propionate molar ratio was lower at lower pH. The amounts and composition of the VFA were not different when using both types of inocula. Thus, we believe that micro-organisms colonising plant fibres obviously do not play an important role in inulin splitting.

Multinucleated giant cell neoplasm of pancreas: light and electron microscopy features.

The morphological features, as demonstrated by electron microscopy, of a rare pancreatic neoplasm indistinguishable from giant cell tumor of bone by light microscopy is presented. These findings are compared with those of the only other case studied by electron microscopy and reported. Features were noted in both studies that are strongly suggestive of the epithelial origin of this neoplasm. However, there were similarities between the material from the current case and giant cell tumor of bone. Degree of differentiation seems to be the most likely determinant of the degree of similarity of the pancreatic neoplasm to the skeletal lesion.