The effect of maternal habitus on macronutrient content of human milk colostrum.

OBJECTIVE: There is a paucity of studies on the impact of maternal body mass index (BMI) on macronutrient content of human milk colostrum (HMC). The objective of this study was to compare macronutrient content of HMC in healthy women of term infants in relation to their BMI. We hypothesized that mother habitus influences human milk colostrum content. METHOD: Colostrum was collected from 109 healthy mothers of hospitalized healthy term infants divided into four prepregnancy BMI groups: 12 underweight, 59 normal weight, 20 overweight, and 18 obese women between 24 and 72 h after birth. Macronutrient content was measured using mid-infrared spectroscopy. RESULTS: There were no significant differences in macronutrients between the BMI groups. We performed four separate stepwise backward multiple regression analyses taking into account fat, carbohydrate, protein or energy content as dependent variables and maternal BMI, parity, gestational age, infant gender, maternal age, maternal education, mode of delivery and time postdelivery. In these analyses, fat, carbohydrate and energy content were not related to maternal BMI, while protein content was significantly and positively correlated with BMI (P=0.008) and negatively correlated with gestational age (P=0.004) and time postdelivery (P<0.001). Colostrum carbohydrate content was positively correlated with parity. Colostrum fat and energy content were negatively correlated with maternal age and positively correlated with parity. CONCLUSION: Most macronutrient and energy content of colostrum are unaffected by prepregnancy maternal BMI, with the exception of protein content that is positively related to maternal BMI.

Disrupted nitric oxide signaling due to GUCY1A3 mutations increases risk for moyamoya disease, achalasia and hypertension.

Moyamoya disease (MMD) is a progressive vasculopathy characterized by occlusion of the terminal portion of the internal carotid arteries and its branches, and the formation of compensatory moyamoya collateral vessels. Homozygous mutations in GUCY1A3 have been reported as a cause of MMD and achalasia. Probands (n = 96) from unrelated families underwent sequencing of GUCY1A3. Functional studies were performed to confirm the pathogenicity of identified GUCY1A3 variants. Two affected individuals from the unrelated families were found to have compound heterozygous mutations in GUCY1A3. MM041 was diagnosed with achalasia at 4 years of age, hypertension and MMD at 18 years of age. MM149 was diagnosed with MMD and hypertension at the age of 20 months. Both individuals carry one allele that is predicted to lead to haploinsufficiency and a second allele that is predicted to produce a mutated protein. Biochemical studies of one of these alleles, GUCY1A3 Cys517Tyr, showed that the mutant protein (a subunit of soluble guanylate cyclase) has a significantly blunted signaling response with exposure to nitric oxide (NO). GUCY1A3 missense and haploinsufficiency mutations disrupt NO signaling leading to MMD and hypertension, with or without achalasia.

Effect of maternal body mass index on cord blood erthropoietin concentrations.

OBJECTIVE: To examine the hypothesis that maternal body mass index (BMI) (an index of maternal adiposity) correlates with cord blood concentrations of erythropoietin (EPO). STUDY DESIGN: Cross-sectional cohort study of consecutively born singleton term healthy infants born to mothers with various BMIs. Excluded were infants with major factors known to be associated with a potential increase in fetal erythropoiesis. Prepregnancy maternal BMI was calculated from maternal recollection. RESULT: There was a significant correlation between EPO concentrations and maternal BMI (R = 0.427, P = 0.007). This correlation remained significant in multiple stepwise regression analysis using the EPO concentration as the dependent variable, and maternal age, parity, gestational age and Apgar scores (1 or 5 min) as potential confounders. CONCLUSION: Cord blood concentrations of EPO correlate with maternal BMI. We speculate that increasing maternal BMI may represent a relative hypoxic burden on the fetus.

Using topical benzocaine before lingual frenotomy did not reduce crying and should be discouraged.

AIM: The US Food and Drug Administration has said that oral preparations containing benzocaine should only be used in infants under strict medical supervision, due to the rare, but potentially fatal, risk of methemoglobinemia. This study aimed to determine the analgesic effect of topical application of benzocaine prior to lingual frenotomy in infants with symptomatic tongue-tie. We hypothesised that the duration of crying immediately following frenotomy with topical benzocaine would be shorter than with no benzocaine. METHODS: This randomised controlled study compared the length of crying after lingual frenotomy in term infants who did, or did not, receive topical application of benzocaine to the lingual frenulum prior to the procedure. RESULTS: We recruited 21 infants to this study. Crying time was less than one minute in all of the subjects. The average length of crying in the benzocaine group (21.6 ± 13.6 sec) was longer than the length of crying in the control group (13.1 ± 4.0 sec), p = 0.103. CONCLUSION: Contrary to our hypothesis, infants who were treated with topical benzocaine did not benefit from topical analgesia in terms of crying time. The use of benzocaine for analgesia prior to lingual frenotomy in term infants should therefore be discouraged.

Isolated mild white matter signal changes in preterm infants: a regional approach for comparison of cranial ultrasound and MRI findings.

OBJECTIVE: To compare echogenicity detected using cranial ultrasound (cUS) and diffuse excessive high signal intensity (DEHSI) detected using magnetic resonance imaging (MRI) by identical region-based scoring criteria in preterm infants. To explore the association between these white matter (WM) signal changes with early neurobehavior. STUDY DESIGN: Forty-nine pre-selected premature infants with only echogenicity on a first routine cUS1 underwent MRI and a repeated cUS2 at term equivalent age. Echogenicity and DEHSI were graded in various brain areas and diffusivity values were calculated. Neurobehavior was assessed using the Rapid Neonatal Neurobehavioral Assessment Procedure. RESULT: WM signal changes were significantly higher on cUS1 than cUS2; and higher in MRI than cUS2 in posterior regions. Infants with DEHSI demonstrated reduced tissue integrity. Imaging findings were not correlated with early neurobehavior. CONCLUSION: Echogenicity and DEHSI likely represent the same phenomenon. Reduction of over-interpretation of WM signal changes may help define criteria for the judicious use of imaging in routine follow-up of premature infants.

Major losses of fat, carbohydrates and energy content of preterm human milk frozen at -80°C.

OBJECTIVE: Long-term storage of human milk (HM) requires freezing at low temperatures, the consequences of which upon macronutrients are unclear. To test the null hypothesis that HM freezing and storage for a range of 1 to 10 weeks at -80 °C does not affect HM fat, protein, lactose and energy contents. STUDY DESIGN: Samples of HM were obtained from 20 mothers (60 samples) of preterm infants (25 to 35 weeks gestation), who routinely expressed their milk, every 3 h, using an electric pump, from the second to the seventh week after delivery. All samples were frozen at -80 °C for 8 to 83 days (43.8 days average). After thawing and homogenization, energy and macronutrient contents were measured using an HM analyzer. RESULT: Fat, carbohydrates and energy contents were significantly lower in thawed HM than in fresh HM (fat, fresh vs thawed: 3.72±1.17 vs 3.36±1.19 g/100 ml, P<0.001; carbohydrates, fresh vs thawed: 5.86±0.71 vs 4.09±0.96 g/100 ml, P<0.001; energy, fresh vs thawed: 64.93±12.97 vs 56.63±16.82 kcal/100 ml, P<0.0001), whereas protein content remained unchanged (protein, fresh vs thawed: 1.14±0.36 vs 1.15±0.37 g/100 ml, P=0.7). The decline in carbohydrates content but not in fat and energy correlated significantly with freezing duration. CONCLUSION: Freezing at -80 °C significantly decreases the energy content of HM, both from fat and carbohydrates. Since quantitatively the decrease in macronutrients was much higher than that published for HM storage at -20 °C, our results do not support freezing HM at -80 °C as the gold standard for long-term storage. We suggest that caloric intake calculations in preterm infants cannot be established based upon fresh HM data.

Conclusiveness of the Cochrane reviews in nutrition: a systematic analysis.

BACKGROUND/OBJECTIVE: To assess the conclusiveness of the Cochrane Reviews (CRs) in the field of Nutrition, we tested the hypotheses that: (1) the majority of CRs is inconclusive; (2) the majority of CRs recognizes the need for further and better studies and (3) the ability to reach a conclusion is dependent on the number of studies performed and number of patients enrolled. SUBJECTS/METHODS: We selected all 87 CRs in the field of Nutrition available in Cochrane library. Each CR was analyzed for the number of randomized clinical trials (RCTs) found, number of RCTs included for analysis, number of patients enrolled, the stated need for further studies and the reason(s) for it and the conclusiveness of the CR. RESULTS: Fifty-six out of eighty-seven CRs (64.4%) were conclusive. The average number of available articles, the percentage of articles included, the average number of RCT's retained in the analyses and the total cumulative number of patients enrolled in the studies retained for analysis were significantly higher in conclusive CRs than in non-conclusive ones. The majority of inconclusive CRs (70.9%) recognized the need for further studies, a percentage not significantly different from that found in conclusive ones (58.9%, P=0.26). The percentage of conclusive CRs was not affected by year of publication. CONCLUSIONS: The majority of CRs in Nutrition is conclusive, but most of them emphasize the need for further studies. The ability for a CR to reach a conclusion is affected by the cumulative patient sample size and number of RCT's included in the analysis.

Quick identification of febrile neonates with low risk for serious bacterial infection: an observational study.

OBJECTIVE: To examine the possible usefulness of simple and quick criteria for identifying febrile neonates with low risk for serious bacterial infection (SBI). DESIGN: All febrile neonates who were admitted between August 1998 and August 2003 to the Pediatric Emergency Department, HaEmek Medical Center, Afula, Israel, and to the Poriya Hospital, Tiberias, Israel, were included in the study. The recommended evaluation of each neonate included details of medical history and a complete physical examination, including blood culture, erythrocyte sedimentation rate (ESR), white cell count (WBC), and analysis and culture of urine and cerebrospinal fluid. Other tests were carried out as necessary. Patients who met all the following criteria were considered to have low risk for SBI: (1) unremarkable medical history; (2) good appearance; (3) no focal physical signs of infection; (4) ESR <30 mm at the end of the first hour; (5) WBC 5000-15 000/mm(3); (6) a normal urine analysis by the dipstick method. RESULTS: Complete data were available for 386 neonates. SBI was documented in 108 (28%) neonates, of whom 14% had a urinary tract infection, 9.3% had acute otitis media, 2.3% had pneumonia, 1.3% had cellulitis, 0.5% had bacterial meningitis and 0.5% had bacterial gastroenteritis. The overall incidence of SBI was 1 in 166 (0.6%) neonates who fulfilled the criteria compared with 107 in 220 (48.6%) in the neonates who did not fulfil the criteria (p<0.001). The negative predictive value for SBI of the combination of the low-risk criteria was 99.4% (95% confidence interval 99.35% to 99.45%). CONCLUSIONS: Fulfillment of the criteria for low risk might be a reliable and useful tool for excluding SBI in febrile neonates.

Expression and regulation of CReMM, a chromodomain helicase-DNA-binding (CHD), in marrow stroma derived osteoprogenitors.

This study follows the expression of CReMM, a new CHD family member, in osteoprogenitors. CReMM expression was analyzed in primary cultured mesnchymal cells from rat and human. Analysis in ex vivo cultured marrow stromal cells (MSC) from rats revealed higher level of CReMM in cells from young (3 months), when compared to cells from old (15 months) rats. CReMM level was higher in human MSC then in mature trabecular bone cells (TBC). Within the MSC population, osteogenic clones showed higher levels of CReMM then non-osteogenic ones. We used bone marrow derived osteogenic cell line (MBA-15) to elaborate on the regulation of CReMM expression in correlation with cell proliferation and co-expression with alkaline phosphatase (ALK). CReMM is highly expressed in proliferating cells and is inversely related to expression of ALK. MBA-15 cells were challenged with dexamethasone (Dex) or 17beta-estradiol and quantification of CReMM at the protein (ELISA) and mRNA (RT-PCR) levels had shown that Dex upregulated CReMM levels. Since CReMM is regulated by Dex, we analyzed the interaction of CReMM with the glucocorticoid receptor (GR), which mediates Dex action. Co-immunopercipitation (Co-IP) demonstrated an association between CReMM and GR. In summary, CReMM is a CHD protein expressed by osteoprogenitors, and we suggest it plays a role in mediating transcriptional response to hormones that coordinate osteoblast function.

Characterization of adhesion and differentiation markers of osteogenic marrow stromal cells.

Marrow stroma cells (MSC) play a major role in osteogenesis. The potential of the MSC to differentiate to bone-forming cells relies upon molecular regulation. This study analyzed MBA-15 cells for the expression of genes and proteins that are key regulators of osteoblast differentiation. These cells express Cbfa1 and c-fos transcription factors (TF) of osteoprogenitor proliferating cells. RT-PCR and immunohistochemistry were used to demonstrate the message and protein expression of extracellular matrix proteins that are a prerequisite for matrix formation and mineralization, including alkaline phosphatase (ALP), osteocalcin, osteopontin, biglycan, and bone sialoprotein (BSP). The activity of ALP was correlated at various cell densities with co-expression of osteocalcin or osteopontin. Adhering cells must attach to the appropriate matrix to enable survival and differentiation. Using attachment assays, we demonstrated that MBA-15 cells adhered to collagenous matrix and the effect on survival measured by changes in intracellular calcium (Ca) levels. The cells' adhesion to matrix is mediated via cell surface molecules. We quantified the expression of cells surface molecules that are important players in mediating cell-matrix interaction. Flow cytometry analysis (FACS) was used to determine the expression of CD-31 (36%), and lower levels were identified for CD-62E and CD11b. In summary, the present study demonstrates the expression of molecular markers that are distinctive for the osteoblastic phenotype in MBA-15 marrow stroma cells and have crucial role in cell-matrix interaction, in establishing the cellular osteogenic phenotype and their survival.

Identification of cultured progenitor cells from human marrow stroma.

The marrow stromal cells (MSC) are essential for regulation of bone remodeling and hematopoiesis. It is of prime importance to isolate MSC and to expand the proliferating cells ex vivo. In this study, we analyzed cultured MSC for various cellular parameters, including cell morphology, cell cycle, and expression of cell surface antigens by flow cytometry. MSC were divided based on cell size to small (S-cells) and large (L-cells) and were visualized by light and electron microscope. The S-cells were proliferating cells correlated with G0/G1 phase of cell cycle, and expressed cFOS. The expression of surface markers CD-34, -44, -51, -61, -62E, -62P, -62L was quantified using flow cytometry. CD-44 was ubiquitously expressed by S and L cells, CD-51 and -61 were expressed by 30%-38% of S-cells. CD-34 and -62 expressed 20% positive of the analyzed cells that were of the proliferating progenitors (S-cells). This study enables the identification of subpopulations from MSC with special attention paid to the proliferating cells from ex vivo cultures of marrow stroma.

Cellular and molecular properties associated with osteosarcoma cells.

Osteosarcoma cells are recognized by abnormal function that causes a primary bone tumor. Osteosarcoma cells U(2)OS and SAOS-2 were analyzed for the expression of cell surface markers. High expression was quantified for hyaloronidase receptor (CD-44) > moderate for integrins (CD-51 and -61), > and lower for selectins (CD-62). High mitotic capacity were demonstrated by gene expression (measured by RT-PCR) and the protein level (measured by FACS) for cFOS, cMYC, and cJUN. The basic definition of osteosarcoma is excessive production of pathological osteoid. Expression of mRNA for matrix genes osteocalcin, osteonectin, and biglycan was studied. Osteocalcin and osteonectin were detected in RNA from primary cultured marrow stromal, trabecular bone cells, and osteosarcoma cell lines (U(2)OS, SAOS-2). mRNA for biglycan was detected only in primary cells and MG-63 cell line and was undetectable in RNA from U(2)OS, SAOS-2 osteosarcoma cell lines and by RNA extracted from bone biopsies of osteosarcoma patients. The absence of biglycan message observed in osteosarcoma samples provides evidence for the alterations in the extra cellular matrix which result with non-mineralized osteoid produced by the osteosarcoma cells.

A history of migraine is not a risk factor to develop an ischemic stroke in the elderly.

OBJECTIVE: To assess the prevalence of migraine in elderly patients hospitalized with ischemic stroke compared with vascular and nonvascular control groups. BACKGROUND: Migraine is a disease with a presumed vascular mechanism. While migraine is a common complaint of young victims of ischemic stroke, it is unclear whether a current or past history of migraine constitutes a risk factor for developing an ischemic stroke in the elderly. METHODS: We obtained current and past headache history from 100 consecutive patients hospitalized with ischemic stroke (aged 60 years or older) and compared the results with 100 patients hospitalized due to acute myocardial infarction and 100 hospitalized patients with no vascular disease. RESULTS: The sex and the age of the patients did not differ among the groups. The lifetime prevalence of migraine (8% of the patients with ischemic stroke, 8% of the nonvascular controls, and 15% of the patients with acute myocardial infarction) or of all types of headaches (27%, 30%, and 15%, respectively), did not differ significantly between the groups. CONCLUSIONS: Based on the reported history, elderly migraineurs are not at increased risk to develop ischemic stroke.

[Thrombocytopenic purpura as sole manifestation of brucellosis in children].

Thrombocytopenic purpura associated with brucellosis has rarely been described in children. The thrombocytopenic purpura is usually part of the array of manifestations of brucellosis, such as fever, malaise, arthralgia, arthritis, hepatosplenomegaly and lymphadenopathy. We describe a 4-year-old girl in whom severe thrombocytopenic purpura was the only manifestation of brucellosis and resolved after appropriate antibiotic therapy. We conclude that brucellosis should be included in the differential diagnosis of thrombocytopenic purpura in areas endemic for brucellosis, and when there is a history of exposure to infected food products.

Normoblasts in large for gestational age infants.

Thirty one term large for gestational age (LGA) infants of non-diabetic mothers were compared with 30 appropriate for gestational age controls. Median absolute nucleated red blood cell counts, lymphocyte counts, and packed cell volumes were significantly higher in the LGA infants than the controls. It is possible that LGA babies of non-diabetic mothers are exposed to relative intrauterine hypoxia.

Plasma fibrinogen in Israeli Moslem and Jewish school-children: distribution and relation to other cardiovascular risk factors. The Petah Tikva project.

Plasma fibrinogen levels were measured in 1,175 Israeli schoolchildren, aged 9-18 years, originating from diverse ethnic groups. The Moslem children displayed striking differences in levels and age-patterns of fibrinogen compared to Jewish children. Lower mean plasma fibrinogen levels in boys aged 9-10, 13-14 and 16-18 were observed among Moslem boys, compared to their Jewish counterparts born in Israel. Moslem girls showed lower levels of fibrinogen than Jewish Israeli girls at ages 9-10 and 16-18. While the Jewish children displayed an age-associated fibrinogen pattern comparable at the three age groups, the levels for Moslem children increased pre-puberty and decreased post-puberty, peaking at 13-14 years. No significant difference in mean plasma fibrinogen was found between sexes within ethnic groups. A number of statistically significant but low correlations (-0.32 to 0.24) were found between plasma fibrinogen and high density lipoprotein cholesterol (HDL-C) as a percentage of total cholesterol, triglycerides, blood glucose, uric acid, blood pressure, Quetelet index and sports activity. The difference between fibrinogen levels might point to a possible ethnicity risk factor explanation rather than environmentally acquired factors.

Divergent age-associated patterns of high density lipoprotein cholesterol and its percentage in Jewish and Moslem Arab Israeli children and adolescents: the Petach Tikva Project.

We studied three groups of Israeli Jewish schoolchildren in and surrounding Petach Tikva, Israel, cross-sectionally, at ages 9-10, 13-14, and 16-18 years, and compared lipid and lipoprotein levels and age-associated lipoprotein patterns in the same age groups of boys and girls in neighboring Israeli Moslem Arab schools during 1986-1987. Moslem children displayed striking differences in the levels of lipids and age-associated patterns of lipoproteins compared with Jewish schoolchildren. The mean total cholesterol levels were lower in the Moslem children, in both sexes, in every age grouping. High density lipoprotein cholesterol (HDL-C) levels were significantly higher at age 16-18 in the Moslem boys than in the Jewish boys. While the Jewish boys displayed a previously reported "typical" pattern of lower HDL-C levels postpuberty compared with prepuberty, the Moslem boys had markedly higher mean HDL-C levels at age 16-18 compared with those at age 9-10. The Moslem girls also had higher HDL-C levels at age 16-18 than those observed in the age 9-10 group. Concomitantly higher HDL-C levels (HDL-C/total cholesterol (%)) were seen in the Moslem boys and girls, at age 16-18 compared with age 9-10, but were not observed in the Jewish children. The identification of an ethnic group in whom HDL-C appears to increase at or near puberty could provide opportunities to elucidate factors that may increase HDL-C in individuals or in populations.