Grandmaternal cells in cord blood.

BACKGROUND: During pregnancy a feto-maternal exchange of cells through the placenta conducts to maternal microchimerism (Mc) in the child and fetal Mc in the mother. Because of this bidirectional traffic of cells, pregnant women have also acquired maternal cells in utero from their mother and could transfer grandmaternal (GdM) cells to their child through the maternal bloodstream during pregnancy. Thus, cord blood (CB) samples could theoretically carry GdMMc. Nevertheless this has never been demonstrated. METHODS: Using Human Leukocyte Antigen (HLA)-specific quantitative PCR assays on three-generation families, we were able to test 28 CB samples from healthy primigravid women for GdMMc in whole blood (WB) and isolated cells (PBMC, T, B, granulocytes, stem cells). FINDINGS: Five CB samples (18%) had GdMMc which could not be confounded with maternal source, with quantities 100 fold lower than maternal Mc in WB and PBMC. Risk of aneuploidies and/or related invasive prenatal procedures significantly correlated with the presence of GdMMc in CB (p=0.024). Significantly decreased HLA compatibility was observed in three-generation families from CB samples carrying GdMMc (p=0.019). INTERPRETATION: Transgenerational transfer of cells could have implications in immunology and evolution. Further analyses will be necessary to evaluate whether GdMMc in CB is a passive or immunologically active transfer and whether invasive prenatal procedures could trigger GdMMc. FUNDING: Provence-Alpes-Côte d'Azur APEX grant # 2012_06549E, 2012_11786F and 2014_03978) and the Foundation for Medical Research (FRM Grant #ING20140129045).

Factors Predicting the Presence of Maternal Cells in Cord Blood and Associated Changes in Immune Cell Composition.

Background: Cord blood (CB) samples are increasingly used as a source of hematopoietic stem cells in transplantation settings. Maternal cells have been detected in CB samples and their presence is associated with a better graft outcome. However, we still do not know what influences the presence of maternal microchimerism (MMc) in CB samples and whether their presence influences CB hematopoietic cell composition. Patients and Methods: Here we test whether genetic, biological, anthropometric and/or obstetrical parameters influence the frequency and/or quantity of maternal Mc in CB samples from 55 healthy primigravid women. Mc was evaluated by targeting non-shared, non-inherited Human Leukocyte Antigen (HLA)-specific real-time quantitative PCR in whole blood and four cell subsets (T, B lymphocytes, granulocytes and/or hematopoietic progenitor cells). Furthermore CB samples were analyzed for their cell composition by flow cytometry and categorized according to their microchimeric status. Results: MMc was present in 55% of CB samples in at least one cell subset or whole blood, with levels reaching up to 0.3% of hematopoietic progenitor cells. Two factors were predictive of the presence of MMc in CB samples: high concentrations of maternal serological Pregnancy-Associated-Protein-A at first trimester of pregnancy (p=0.018) and feto-maternal HLA-A and/or -DR compatibility (p=0.009 and p=0.01 respectively). Finally, CB samples positive for MMc were significantly enriched in CD56+ cells compared to CB negative for MMc. Conclusions: We have identified two factors, measurable at early pregnancy, predicting the presence of maternal cells in CB samples at delivery. We have shown that MMc in CB samples could have an influence on the hematopoietic composition of fetal cells. CD56 is the phenotypic marker of natural killer cells (NK) and NK cells are known to be the main effector for graft versus leukemia reactions early after hematopoietic stem cell transplantation. These results emphasize the importance of MMc investigation for CB banking strategies.

Soluble HLA-G Expression Inversely Correlates With Fetal Microchimerism Levels in Peripheral Blood From Women With Scleroderma.

Women with scleroderma (SSc) maintain significantly higher quantities of persisting fetal microchimerism (FMc) from complete or incomplete pregnancies in their peripheral blood compared to healthy women. The non-classical class-I human leukocyte antigen (HLA) molecule HLA-G plays a pivotal role for the implantation and maintenance of pregnancy and has often been investigated in offspring from women with pregnancy complications. However data show that maternal HLA-G polymorphisms as well as maternal soluble HLA-G (sHLA-G) expression could influence pregnancy outcome. Here, we aimed to investigate the underlying role of maternal sHLA-G expression and HLA-G polymorphisms on the persistence of FMc. We measured sHLA-G levels by enzyme linked immunosorbent assay in plasma samples from 88 healthy women and 74 women with SSc. Male Mc was quantified by DYS14 real-time PCR in blood samples from 58 women who had previously given birth to at least one male child. Furthermore, eight HLA-G 5'URR/3'UTR polymorphisms, previously described as influencing HLA-G expression, were performed on DNA samples from 96 healthy women and 106 women with SSc. Peripheral sHLA-G was at lower concentration in plasma from SSc (76.2 ± 48.3 IU/mL) compared to healthy women (117.5 ± 60.1 IU/mL, p < 0.0001), independently of clinical subtypes, autoantibody profiles, disease duration, or treatments. Moreover, sHLA-G levels were inversely correlated to FMc quantities (Spearman correlation, p < 0.01). Finally, women with SSc had lower sHLA-G independently of the eight HLA-G 5'URR/3'UTR polymorphisms, although they were statistically more often homozygous than heterozygous for HLA-G polymorphism genotypes -716 (G/T), -201 (G/A), 14 bp (ins/del), and +3,142 (G/A) than healthy women. In conclusion, women with SSc display less sHLA-G expression independently of the eight HLA-G polymorphisms tested. This decreased production correlates with higher quantities of persisting FMc commonly observed in blood from SSc women. These results shed some lights on the contribution of the maternal HLA-G protein to long-term persistent fetal Mc and initiate new perspectives in this field.

Loss of caspase-8 protects mice against inflammation-related hepatocarcinogenesis but induces non-apoptotic liver injury.

BACKGROUND & AIMS: Disruption of the nuclear factor-κB (NF-κB) essential modulator (NEMO) in hepatocytes of mice (NEMO(Δhepa) mice) results in spontaneous liver apoptosis and chronic liver disease involving inflammation, steatosis, fibrosis, and development of hepatocellular carcinoma. Activation of caspase-8 (Casp8) initiates death receptor-mediated apoptosis. We investigated the pathogenic role of this protease in NEMO(Δhepa) mice or after induction of acute liver injury. METHODS: We created mice with conditional deletion of Casp8 in hepatocytes (Casp8(Δhepa)) and Casp8(Δhepa)NEMO(Δhepa) double knockout mice. Acute liver injury was induced by Fas-activating antibodies, lipopolysaccharides, or concanavalin A. Spontaneous hepatocarcinogenesis was monitored by magnetic resonance imaging. RESULTS: Hepatocyte-specific deletion of Casp8 protected mice from induction of apoptosis and liver injury by Fas or lipopolysaccharides but increased necrotic damage and reduced survival times of mice given concanavalin A. Casp8(Δhepa)NEMO(Δhepa) mice were protected against steatosis and hepatocarcinogenesis but had a separate, spontaneous phenotype that included massive liver necrosis, cholestasis, and biliary lesions. The common mechanism by which inactivation of Casp8 induces liver necrosis in both injury models involves the formation of protein complexes that included the adaptor protein Fas-associated protein with death domain and the kinases receptor-interacting protein (RIP) 1 and RIP3-these have been shown to be required for programmed necrosis. We demonstrated that hepatic RIP1 was proteolytically cleaved by Casp8, whereas Casp8 inhibition resulted in accumulation of RIP complexes and subsequent liver necrosis. CONCLUSIONS: Inhibition of Casp8 protects mice from hepatocarcinogenesis following chronic liver injury mediated by apoptosis of hepatocytes but can activate RIP-mediated necrosis in an inflammatory environment.

Activation of ectopic Oct-4 and Rex-1 promoters in human amniotic fluid cells.

Recently, amniotic fluid was suggested as a new source for stem-cell research and tissue engineering approaches. In order to enable isolation of stem cells and establishment of lines of such cells with an undifferentiated phenotype we have introduced green fluorescent protein regulated by the promoters of the stem cell-specific genes, Oct-4 or Rex-1, into human amniotic fluid cells. For the introduction of DNA into human amniotic fluid cells, we have optimized a specific transfection protocol. We found that human amniotic fluid contains cell populations which are able to activate these promoters. These undifferentiated cells expressing green fluorescent protein can be analysed on a flow cytometer. In addition, we have introduced a plasmid harboring a neomycin-resistance gene under the control of the Oct-4 promoter. G418 selection allowed the isolation of undifferentiated stem cells expressing Oct-4 protein out of human amniotic fluid samples. Our findings confirm the existence of stem cells within amniotic fluid. In addition, the ability to transfect human amniotic fluid cells and to isolate stem-cell marker-positive cells will provide the means to study and manipulate these cells for the purpose of basic and applied research.

Escherichia coli: the best biological drinking water indicator for public health protection.

Public health protection requires an indicator of fecal pollution. It is not necessary to analyse drinking water for all pathogens. Escherichia coli is found in all mammal faeces at concentrations of 10 log 9(-1), but it does not multiply appreciably in the environment. In the 1890s, it was chosen as the biological indicator of water treatment safety. Because of method deficiencies, E. coli surrogates such as the 'fecal coliform' and total coliforms tests were developed and became part of drinking water regulations. With the advent of the Defined Substrate Technology in the late 1980s, it became possible to analyse drinking water directly for E. coli (and, simultaneously, total coliforms) inexpensively and simply. Accordingly, E. coli was re-inserted in the drinking water regulations. E. coli survives in drinking water for between 4 and 12 weeks, depending on environmental conditions (temperature, microflora, etc.). Bacteria and viruses are approximately equally oxidant-sensitive, but parasites are less so. Under the conditions in distribution systems, E. coli will be much more long-lived. Therefore, under most circumstances it is possible to design a monitoring program that permits public health protection at a modest cost. Drinking water regulations currently require infrequent monitoring which may not adequately detect intermittent contamination events; however, it is cost-effective to markedly increase testing with E. coli to better protect the public's health. Comparison with other practical candidate fecal indicators shows that E. coli is far superior overall.

Outcomes of a nursing intervention for siblings of chronically ill children: a pilot study.

PURPOSE: To evaluate the outcomes of a structured, educational, and support group intervention (ISEE, Intervention for Siblings: Experience Enhancement) for siblings of children with chronic illness (cancer, cystic fibrosis, diabetes, and spina bifida), including a session with parents about sibling needs; and to describe sibling and parent perceptions of sibling experiences at home. DESIGN: One-group, pretest-posttest pilot study. PARTICIPANTS: A convenience sample of 22 siblings and parents. SETTING: A Midwestern university medical center. MAIN OUTCOME MEASURES: Knowledge of Illness Test, parent ratings on a global, single item, 10-point scale. RESULTS: Sibling test scores increased significantly after intervention, compared to baseline. Parents' average evaluation rating was 9 on a 10-point scale. Parents supported their positive ratings with verbatim descriptions. Sibling and parent perceptions of sibling experiences were congruent, suggesting the sources of potential adjustment problems in siblings, and were consistent with the literature. CONCLUSIONS: A randomized, clinical trial with a larger sample size is needed to evaluate the intervention further.

Illusory safeguards: legitimizing distortion in recall with guidelines for forensic hypnosis--two case reports.

Two amnesic automobile accident victims remembered the information needed for their ongoing lawsuits during hypnosis. Meeting the recording requirements of the Hurd safeguards led to the admission of hypnotically influenced testimony in court in one case, whereas failure to record led to exclusion in the other. In both cases, closed-head trauma almost certainly prevented long-term memory consolidation. Thus adherence to guidelines for forensic hypnosis legitimized distortions in recall instead of preventing them. Hypnosis used to facilitate hypermnesia alters expectations about what can be remembered, makes memory more vulnerable to postevent information, and increases confidence without a corresponding increase in accuracy. Distortion of recall is an inherent problem with the use of hypnosis and hypnotic-like procedures and cannot be adequately prevented by any set of guidelines.

Small bowel obstruction: a morbid complication after laparoscopic herniorrhaphy.

More than 500,000 inguinal herniorrhapies are performed each year in the United States. The traditional methods of hernia repair have demonstrated a low complication rate (< 1%). Recently, the success of laparoscopic cholecystectomy (complication rate < 2%) has generated enthusiasm for the application of laparoscopic techniques toward the management of other intra-abdominal pathologies. The initial reports concerning the efficacy and advantages of laparoscopic surgery are encouraging. However, the limited data available thus far precludes the answering of questions regarding perioperative morbidity. We present two cases of small bowel obstruction that occurred following uncomplicated laparoscopic herniorrhaphy. One occurred 2 weeks postoperatively and consisted of a herniation through the re-approximated peritoneum over the repair site. The second occurred 3 days postoperatively and was caused by herniation through a large (10 mm) trochar site. Surgical intervention involved exploratory laparotomy in the former and laparoscopic techniques in the latter. We suggest that the principles of solid abdominal closure are applicable to the relatively small peritoneal defects created during laparoscopic procedures. Additionally, awareness of this potential complication will aid in decreasing the morbidity of this and other laparoscopic surgeries.

Health risks from contaminated water: do class and race matter?

The impact of contaminants in water on minorities and economically disadvantaged persons was reviewed. Environmental legislation governing water was summarized as background information against which relevant studies were evaluated. The majority of the available information was anecdotal or case study and did not lend itself to making quantitative comparisons or analyses. However, the data did present certain trends that led to the conclusion that inequities concerning exposure to contaminants in water may exist. The following recommendations were made: current data bases should be analyzed and new data bases created to facilitate assessments of exposure to waterborne contaminants to all populations; an analysis of populations not covered by the Safe Drinking Water Act should be undertaken; a survey should be conducted of the drinking water infrastructure and the results evaluated to identify any impacts to minorities and economically disadvantaged persons; the social, cultural and economic characteristics that influence human exposure to waterborne contaminants need to be identified; and better educational and community outreach programs need to be developed and implemented.

Paleoearthquakes in the puget sound region recorded in sediments from lake washington, u.s.a.

Holocene sediments in Lake Washington contain a series of turbidites that were episodically deposited throughout the lake. The magnetic signatures of these terrigenous layers are temporally and areally correlatable. Large earthquakes appear to have triggered slumping on the steep basin walls and landslides in the drainage area, resulting in turbidite deposition. One prominent turbidite appears to have been deposited about 1100 years ago as the result of a large earthquake. Downcore susceptibility patterns suggest that near-simultaneous slumping occurred in at least three separate locations, two of which now contain submerged forests. Several other large earthquakes may have occurred in the last 3000 years.

Hypnotic susceptibility and its relationship to outcome in the behavioral treatment of asthma: some preliminary data.

12 subjects from an experiment on relaxation therapy for asthma were given the Harvard Group Scale of Hypnotic Susceptibility, Form A. Full scale hypnotic susceptibility scores were positively correlated, at a borderline significance, with improvement in the methacholine challenge test, a measure of asthma severity. Performance on the amnesia item of the Harvard Group Scale was correlated with improvement in self-reported symptoms of asthma.

Waterborne disease in Colorado: three years' surveillance and 18 outbreaks.

The Colorado Department of Health conducted intensive surveillance for waterborne diseases during the three-year period July 1, 1980-June 30, 1983. Eighteen outbreaks of waterborne illness were investigated. Outbreaks involved from 15 to 1,500 ill persons. Giardia lamblia was confirmed or suspected as the agent in nine outbreaks, rotavirus in one, and no agent could be identified in eight. Seventeen outbreaks occurred on surface-water systems; none of these had adequate chemical pretreatment and filtration. Investigation of water systems exhibiting positive coliform results during the first year detected no outbreaks. Activities important to effective surveillance included educational outreach programs to local health agencies, physicians and the public, and the designation of one individual to whom all water-related public complaints and health department inquiries were directed.

Septic cavernous sinus thrombosis.

Septic cavernous sinus thrombosis remains a potentially lethal disease, although advances in therapy have lowered the morbidity and mortality substantially. Prevention, early diagnosis, and aggressive treatment offer the best hope for recovery.

A community waterborne gastroenteritis outbreak: evidence for rotavirus as the agent.

A community waterborne nonbacterial gastroenteritis outbreak occurred in Eagle-Vail, Colorado in March 1981. Illness (defined as vomiting and/or diarrhea) was statistically associated with water consumption (chi 2 for linear trend = 7.07, p less than .005). Five of seven persons associated with the outbreak were infected with rotavirus as shown by virus detection or serological methods. Bacterial pathogens, Giardia lamblia, and Norwalk virus were excluded as responsible agents. Rotavirus should be looked for as a cause of waterborne outbreaks.

Extracranial carotid artery injury. Current surgical management.

Fifty patients were treated for extracranial carotid artery trauma during the period from 1954 through 1981. The overall mortality rate was 20 percent. Primary repair was achieved in 38 patients (76 percent) with a mortality of 7.8 percent, and ligation was performed in six patients (12 percent) with a 50 percent mortality. The left carotid system was injured more frequently than the right; hemorrhage from an isolated carotid injury was readily controlled by digital pressure. Associated injuries affected the prognosis adversely. Significant neurologic deficits were present in 12 patients on first admission but the presence of neurologic deficit or coma did not serve as a contraindication to arterial repair and restoration of cerebrovascular continuity. Arteriography was not utilized once the clinical diagnosis of carotid arterial injury was made, but it was utilized when the proximity of the cervical injury provided the only suggestive evidence that vascular damage may have occurred. The presence of significant shock, coma, or neurologic deficit represent adverse prognostic features but do not contraindicate carotid repair. Profuse bleeding from a carotid injury or the presence of severe posthemorrhagic shock require urgent surgery. If control of bleeding is difficult to achieve, then ligation is preferred to suture repair.

Screening for hypertension in the emergency department.

Review of emergency department charts at three university-affiliated hospitals showed that less than half of all patients who had elevated blood pressure (BP) recorded were recognized by physicians to be hypertensive. At the primary teaching hospital, less than one third of patients with the greatest BP elevation (greater than 20 mm Hg above normal) were sent for some type of hypertension follow-up care.