Newborn Specimens Research Articles

Iron stores in maternal and cord blood.

AbstractSerum ferritin concentration was measured in 206 paired maternal and newborn (Umbilical cord sample) blood specimens taken during delivery. Iron deficiency (assessed by serum ferritin assay) was found in 38% pregnant women. The mean serum ferritin was significantly low in pregnant mothers compared to non‐pregnant women, indicating depleted iron stores at term.Serum ferritin levels in cord blood were higher than those in the maternal blood, indicating that the mother is in a state of iron deficiency with low serum ferritin and the fetus in a state of iron sufficiency with high serum ferritin.Maternal ferritin had no correlation with cord ferritin and babies born to iron deficient mothers had low but not statistically significant serum ferritin levels. This suggests that iron storage in the mother and iron metabolism in the fetus are not directly related. However, there seemed to be a level of maternal iron stores below which the newborn was endowed with decreased iron stores.

A new method for the detection of phospholipase A 2 variants: Identification of isozymes in the venoms of newborn and adult Bothrops asper (terciopelo) snakes

E. Moreno, A. Alape, M. Sánchez and J. M. Gutiérrez. A new method for the detection of phospholipase A 2 variants: identification of isozymes in the venoms of newborn and adult Bothrops asper (terciopelo) snakes. Toxicon 26, 363 – 371, 1988. — A new method for the identification of phospholipase A 2 isozymes in snake venoms is described. The technique is based on the separation of the venom components by isoelectric focusing in agarose gels, transfer of the protein bands by diffusion onto nitrocellulose paper and detection of the phospholipolytic activity of the enzymes by a hemolytic assay either in agarose gels or by benzidine reaction on a solid matrix. Striking differences in the electrophoretic patterns of the phospholipase A 2 isozymes between the Atlantic and Pacific venoms and between the newborn and adult venoms from Bothrops asper specimens were observed. The method allowed the detection of 9 different phospholipase A 2 isozymes in the venom of adult Atlantic, 7 isozymes in the venom of adult Pacific, and 2 – 3 isozymes in the venoms of newborn specimens. Horse polyvalent antivenom varied in its capacity to neutralize the phospholipolytic activity of the different isozymes in the same venom and among different venoms.

Detection of proteins antigenically related to Bothrops asper myotoxin in crotaline snake venoms

B. Lomonte, E. Moreno and J. M. Gutiérrez. Detection of proteins antigenically related to Bothrops asper myotoxin in crotaline snake venoms. Toxicon 25, 947 – 955, 1987. — The presence of components antigenically related to Bothrops asper myotoxin was investigated by Western blotting and immunoelectrophoretic techniques. B. asper myotoxin is a non-glycosylated monomeric phospholipase A with a molecular weight by SDS-PAGE of 16,000 and isoelectric point of pH 9.8 – 10.0. Results showed that proteins in the venoms of B. nummifer, B. godmani, B. schlegelii, B. picadoi, and Agkistrodon bilineatus were recognized by monospecific antibodies to B. asper myotoxin raised in rabbit and sheep. Western blotting indicated that cross-reacting proteins have a molecular weight of 16,000, with the exception of that of B. picadoi, which is of 24,000 mol. wt. However, immunoelectrophoresis indicated that these components are highly heterogeneous in charge, ranging from basic to acidic proteins. The cross-reacting component(s) present in newborn B. asper venom has a different charge from that of the ‘adult-type’. Venoms from newborn specimens showed an additional cross-reacting band of 18,000 mol. wt. Myotoxin is an abundant component in adult B. asper venom. Myotoxin - antimyotoxin complexes had different electrophoretic mobilities in rocket immunoelectrophoresis depending upon the species in which monospecific immune sera were produced.

Stability of amino acids and galactose in the newborn screening filter paper blood specimen

A POTENTIAL ADVANTAGE of filter paper blood specimens for neonatal screening of inborn errors of metabolism is the ease with which they can be saved for future studies. The specimen cards may be stored in files or boxes without the requirements of liquid specimens for space, protection against breakage, and refrigeration. Filter paper specimens saved are suitable for certain retrospective metabolic studies, ~'2 but this suitability has not been established for the amino acids that are central to the metabolic disorders for which screening is conducted, or for galactose, also a key metabolite in newborn screening. Information about the stability of metabolites in stored filter paper blood specimens is also necessary when retesting the original specimen to determine whether a child with a metabolic disorder not detected by newborn screening had the abnormality in the newborn period. If the results on retesting are a valid reflection of the neonatal value, biologic variation can be distinguished from error in the testing process~; otherwise, the results on retesting may be misleading. We examined the stability to bacterial assay of certain amino acids and of galactose in filter paper blood specimens by comparing values obtained after years of storage with those recorded when the specimens were initially tested.

Isolation from a polyvalent antivenom of antibodies to a myotoxin in Bothrops asper snake venom

Antibodies against Bothrops asper myotoxin were purified from a polyvalent antivenom by affinity chromatography. These antibodies neutralized most of the myotoxic activity of crude B. asper venom, as judged by histologic evaluation of skeletal muscle and determination of plasma creatine kinase levels in mice. When tested by immunoelectrophoresis, purified antibodies formed two superimposed bands of precipitation against an homogenous (by SDS-PAGE analysis) preparation of B. asper myotoxin, as well as against crude B. asper venom. Ouchterlony immunodiffusion analysis of purified antibodies showed two precipitation bands with a pattern of complete immunologic identity between samples of crude B. asper venoms from specimens collected in the Atlantic and Pacific regions of Costa Rica. In addition, these antibodies precipitated when reacted against venom of newborn B. asper specimens from the Pacific region, but not against venom of newborn specimens from the Atlantic region. Purified antibodies were tested by immunodiffusion against eleven different snake venoms from Costa Rica. Only the venom of B. schlegelii cross-reacted, indicating the presence in this venom of components immunologically related to B. asper myotoxin. Analysis of purified antibodies to B. asper myotoxin by agarose electrophoresis and by SDS-PAGE suggests the presence of both IgG and IgM on the basis of electrophoretic position and molecular weight of the bands. Results obtained in neutralization experiments suggest that this myotoxin is a major factor in the development of local myonecrosis induced by crude B. asper venom.

Selective Cultivation of Human Melanocytes from Newborn and Adult Epidermis

Development of adequate culture systems for the human epidermal melanocyte is critical to further advances in pigment cell biology. We now report selective growth and long-term maintenance of melanocytes derived from both newborn and adult skin specimens. Disaggregated epidermal cell suspensions were plated in a hormone-supplemented medium containing cholera toxin and a hypothalamic extract treated to remove keratinocyte growth-promoting activity. After 3-4 weeks, pure melanocyte populations could be harvested and serially passaged up to 6 times over several months for a total of 10 or more cumulative population doublings in vitro. Electron microscopic studies revealed metabolically active cells with abundant melanosomes in various stages of melanization throughout the culture lifespan. Differences in size and number of melanosomes attributable to race of the tissue donor were readily apparent, and pigment content of melanocytes from both black and Caucasian donors appeared to increase with time in culture. Newborn melanocytes proliferated more rapidly and survived longer than did adult melanocytes, but there were no consistent morphologic differences as a function of donor age. Comparison of growth potential for the 3 major skin-derived cell types in this hormone-supplemented medium revealed striking specificity for melanocytes, with total elimination of keratinocytes over 1-2 weeks, and no fibroblast proliferation whatever in the absence of serum supplementation. This system promises to facilitate in vitro investigation of epidermal melanocytes in normal and diseased human skin.

Virus Replication and Induction of Interferon in Human Epidermal Keratinocytes Following Infection with Herpes Simplex Virus

Keratinocyte cultures derived from surgical skin specimens of healthy newborns and adults were infected with herpes simplex virus (HSV) type 1 or 2. Typical HSV cytopathic effects involved all cell layers in stratified colonies, and paralleled the production of infectious virus. Virus growth curves and production of virus were comparable in newborn and adult keratinocytes. Interferon (IF) production by keratinocytes paralleled the yield of virus over at least 72 h, and was greater in cultures of adult cells than cultures from newborns. UV irradiation of HSV resulted in progressive virus inactivation and a parallel reduction in induced IF. This suggests that IF production was related to virus replication, and that irradiated (noninfectious) HSV DNA did not contribute significantly to the generation of IF in this system. These results establish that human epidermal keratinocytes can serve as a model system for quantitative assessment of herpes simplex virus infection.

Ultrastructural changes in neonatal liver tissue: effects of maternal drinking.

There is already sufficient evidence in the literature that alcohol abuse during pregnancy has a toxic effect upon the developing fetus; however, previous studies have not revealed any morphological changes in fetal or newborn liver specimens from animals exposed to alcohol in utero. As it is known that alcohol freely crosses the placental barrier, this investigation was an attempt to demonstrate that structural abnormalities can indeed be identified in neonatal mouse liver specimens from pups exposed to alcohol in utero. Chosen as a model for this study was the C57BL/KsJ mouse strain as this particular animal demonstrates an alcohol preference paralleling that of the human alcoholic. Findings appear to indicate the presence of abnormal changes on the morphological level in these study animals.

Estudio comparativo de los venenos de serpiente Cascabel ( Crotalus durissus durissus) de ejemplares adultos y recien nacidos

B. Lomonte, J. A. Gené, J. M. Gutiérrez and L. Cerdas. Estudio comparativo de los venenos de serpiente Cascabel ( Crotalus durissus durissus) de ejemplares adultos y recién nacidos. Toxicon 21, 379 – 384, 1983. — Venoms from adult and newborn Central American rattlesnakes ( Crotalus durissus durissus) were compared for lethal, proteolytic, hemorrhagic, myonecrotic, edematigenous and in vitro hemolytic activities. Electrophoretic and immunoelectrophoretic patterns showed some differences between these venoms. Venom from newborn snakes was devoid of hemorrhagic and edematigenous activities, whereas the venom from adult specimens induced these effects. On the other hand, newborn snake venom showed higher lethality and indirect hemolytic activity, and lower proteolytic activity, than venom from adult specimens. Both types of venoms induced only slight myonecrosis in mice, as judged by histological observation. The ed 50 of an antivenom, in terms of absolute weight neutralized per ml of serum, was lower for the newborn specimens venom than for adult's venom, however, for each venom the number of ld 50 neutralized was similar.

Comparison of Primary TSH (pTSH) to Primary T4 with Confirmatory TSH (T4/TSH) Screening for Congenital Hypothyroidism (CH)

Our recent survey of 38 states in U.S. & 10 countries indicated that 4.45 million(m) infants were screened for CH by 12/31/79, & 1048 cases of primary hypothyroidism detected (incidence of 1 in 4245) on the 1st specimen. Screening methods varied: (1) T4/TSH: 3.73m, 835 cases, 1 in 4466 incidence; (2) pTSH: 0.5m, 157 cases (1 in 3239); (3) both T4 & TSH: 0.2m, 56 cases (1 in 3695). In summary, programs using pTSH screening detected a higher incidence of CH than programs using T4/TSH screening. When confirmatory TSH is tested on the lowest 10-15% of T4 values, the incidence of CH approaches a maximum comparable to pTSH screening. In programs collecting 2 routine specimens, 169 cases were detected in 0.78m newborn (nb) specimens (1 in 4600) & 22 cases in 0.6m followup(fu) specimens (1 in 26,000). Of the 38 cases of CH that were missed, 12 had normal T4 levels & 4 with primary CH had normal TSH levels. For 3 years we have tested both nb & fu specimens by 2 methods: (1)T4/TSH (TSH test on lowest 3% of T4) & (2) pTSH. Normal TSH is <30 μU/ml of serum in two 3 mm blood discs. 60% of nb specimens were obtained by age 2 days, & the false positive (elevated TSH) rate was 0.9% whereas the rate in nb specimens collected after 2 days & in all fu speicmens was <0.2%, comparable to pTSH cord serum screening. Summary: For specimens collected before 48h of age, higher TSH cutoff levels(30-60μU/ml)must be determined to avoid false positive recall rates,yet identify all infants with CH.

Urine screening for metabolic disease in newborn infants

A new method for urine screening for metabolic disease in newborn infants is described. A battery of bacterial inhibition assays to test urine-impregnated filter paper from 3- to 4-week-old infants for amino acids, purines, and pyrimidines was used. We were able to establish the accuracy and efficiency of the method by examining 289 unknown specimens from other laboratories and by collecting and testing 18,400 newborn infants' urine specimens. The major advantages over existing chromatography methods are that: (1) the technology to perform the test already exists in most laboratories screening for metabolic disorders; (2) it is relatively inexpensive; (3) collection of the sample is easy and cooperation of the parents is good; (4) the false positive rate is low (0.9%); and (5) tests can be targeted to detect clinically significant disorders. In this screening program, we detected two cases of persistent neonatal tyrosinemia, two cases of cystinuria, and one case of citrullinemia. These results suggest that urine screening is a good adjunct to blood screening of newborn infants.

Evaluation by serum ferritin assay of the influence of maternal iron stores on the iron status of newborns and infants.

Serum ferritin concentration was measured in 47 paired maternal and newborn (umbilical cord sample) blood specimens during delivery. The serum ferritin concentration was also measured in 31 of the above babies at the age of six months. Serum ferritin concentration of the newborn babies (mean 222 microgram/l) born to the mothers with a low serum ferritin content at term (mean 28 microgram/l) was significantly lower (p < 0.05) than that of those (mean 324 microgram/l) born to the mothers with a normal ferritin concentration (mean 102 microgram/l). The difference between the two groups of infants persisted for at least the whole study period of 6 months (mean at 6 months of age 99 microgram/l and 150 microgram/l, respectively).

Simple Modification of the Cellulose Acetate Method for Identifying Adult Hemoglobin (Hemoglobin A) in Dried Specimens of Newborn Blood

Definition of hemoglobin A in cellulose acetate electrophoretograms of eluate from air-dried specimens of newborns' blood was improved by adding glycerine to the Tris--ethylenediametetraacetic acid--borate buffer system. Examination of 1,672 randomly received samples showed no reduction in ability to detect hemoglobin S with the modified buffer. The method may be adopted as a useful adjunct to current technics for rapid electrophoretic screening of neonatal dried blood samples.

Newborn screening for galactosemia and other galactose metabolic defects

Summary Routine newborn screening is necessary for the effective diagnosis of galactosemia. The criticism that routine screening for galactosemia is unnecessary is based on the misconception that the diagnosis will almost always be made clinically, since the disease produces clinical manifestations in the infant. Acutally most newborn infants, detected by routine screening, were not suspected to have galactosemia, even when ill. Furthermore, some infants with clinically important transferase variants are not ill as neonates. In Massachusetts, a black infant who probably has the Negro variant form of galactosemia and an infant with the Rennes variant of galactosemia 6 were clinically normal at five days of age when detected by routine screening. Complications of galactosemia can occur in both variants if treatment is not given. In addition, galactokinase deficiency, which results in cataracts but not in acute illness, 32 cannot be detected on a clinical basis until the cataracts have become apparent, when it is too late for preventive therapy. The full impact of galactosemia on the newborn infant has become evident only since routine newborn screening has been initiated. It is now apparent that death associated with bacterial sepsis may occur in about 30% of those with untreated classical galactosemia. 4 In most of these deaths diagnosis and treatment were delayed until the second week of life. Consequently, screening for galactosemia should be performed on a blood specimen that is obtained no later than the third or fourth day of life and is delivered promptly to the screening laboratory. There should be sufficient medical back-up available to the laboratory so that the attending physician can be quickly contacted by telephone when an abnormality suggesting galactosemia is encountered. Further procedures should follow: (1) Urine should be tested for reducing substance. (2) If urinary reducing substance is present, galactosemia should be presumed; milk feedings should be discontinued; and blood and urine specimens should be sent to a laboratory for confirmatory testing. (3) If the infant is ill, bacterial cultures should be obtained and treatment for sepsis initiated. (4) If galactosemia is confirmed, treatment should be continued. Otherwise, treatment can be discontinued. Galactosemia screening should be routine for all newborn infants. It is a disorder with definite and severe complications, but one in which the complications can be prevented with simple and inexpensive treatment. When a test for galactose and gal-l-P, such as the Paigen assay, is also applied to the newborn blood specimen that is, submitted for PKU screening, galactosemia can be detected efficiently and effectively.

Studies on fluoride distribution in infants and small children.

The fluoride (F) concentrations in hard tissues and blood plasma were determined in biopsy and autopsy specimens from fetuses, infants, and children on varying F intake. In newborn autopsy cases specimens consisting of rib, jawbone with teeth, and blood plasma the F concentrations of bone and dentin were of the same order, those of the enamel lower. Correlations between individual F values in different tissues were all positive, highest for enamel/dentin. Low birth weight entailed only slightly reduced F contents, high water F significantly increased F contents. In the biopsy material (rib and blood plasma) the maximal rib/ash concentration was about 400 parts/10(6) F. A negative correlation between rib F and plasma F is specially commented on. The possible influence of F on bone development as expressed by blood plasma F and phosphatase activity was investigated in three groups of infants aged 2-6 months, either breast-fed or fed dry-milk formulas diluted with drinking waters of different F content. The children of these groups were calculated to ingest F in the ratio of about 1:10:50. The highest F group had higher plasma F values than the two other groups, whereas its alkaline phosphatase was significantly higher than that of the lowest F group only. The influence of disturbances in the skeletal development on F retention studied in physically handicapped children aged 4-15 years living in a city with about 1 part/10(6) F in the drinking water. The severely handicapped showed a higher urinary F excretion than the controls.

The role of basicranial synchondroses in flexure processes and ontogenetic development of the skull base.

AbstractThe contribution of the basicranial synchondroses in the growth of neurocranial length and ontogenetic development of the cranial base were investigated. The study concentrated on the midsphenoidal synchondrosis and its delayed fusion in nonhuman primates when compared to man, and on the spheno‐occipital synchondrosis. The mode and time of fusion of both growth centers were observed, and their role in the ontogenetic growth changes (flattening processes) of the cranial base were established. The chondrogenic ossification of midsphenoidal and spheno‐occipital synchondroses was studied on 20 skulls of Macaca mulatta females, ranging in age from newborn specimens to those 24 months old. The technique of in vivo tetracycline bone labeling was used for histologic evaluation of the material. Different chondrogenic growth patterns were observed in both synchondroses. The endochondral activity of the spheno‐occipital synchondrosis increased with age, from a nonactive narrow cartilaginous column in the neonatal specimen to a broad band with high chondrogenic ossification in the 24‐month‐old specimens. This growth center contributes to elongation of the posterior portion of the cranial base and is a secondary factor in its flexion.The midsphenoidal synchondrosis seems to be the primary factor in the mode of flexure of the cranial base in Macaques. This growth center is very active in the first ten months of life but later exhibits cessation of chondrogenic activity and long remains unfused. The first signs of fusion were observed as late as 72 months of age. At the same time, the continuous process of cranial base flattening showed the first signs of tapering off.

Cyclic Changes in the Structure of Nucleolus-Associated Chromatin in Neurons of Birds

The results of morphological investigations on nucleolus-associated chromatin or central nervous system neurons of new-born and adult specimens of Gallus domesticus are described. Various morphological arrangements of neuronal nucleolus-associated chromatin, related to different degrees of development of the nucleolus are classified into four different types.

The Pre-Natal Development of Skin and hair in Cattle (Bos Taurus L.)

A histological study has been made of the development of the skin and hair follicles in 33 foetuses, ranging in age from 68 to 274 days, and seven new-born specimens, of various breeds of Bos taurus L. Skin samples were taken from the mid-lateral region of the trunk, and both serial sections and whole mount preparations have been examined. Extensive use has been made of' counts of the follicle and hair population as well as skin and hair follicle measurements which hitherto have apparently received little attention in studies of this type.

A Newborn Piked Whale in Tasmania

On June 7, 1957, we examined a small whalebone whale which had been found on the shore at Lewisham in southeastern Tasmania. The animal was much decomposed internally and in the region of the mouth and from its condition we judged it to have been dead at least about a month. Mr. E. C. Clark, who drew our attention to it, said that it had been seen first two weeks before and at that time had the appearance of having been dead for some weeks. We concluded that it had died probably four to six weeks previously and was already dead when it reached the shore at Lewisham. Its very small size—it was only 7 ft. 1 in. (216 cms.) in straight line length—immediately indicated that it was a fetal or newborn specimen. An inch or two of umbilical cord was still visible, although much …

Duplicate Twins and Double Monsters.

Northampton, Mass., Dec. 15, 1904. <h3>To the Editor:</h3> —As I have recently become interested in a line of investigation which requires the careful examination of all forms of double monsters, I would be extremely grateful for the gift or loan of fetal or new-born specimens exhibiting any grade of this phenomenon. As such cases are rare, there is a natural tendency to store them up as museum specimens, in which condition the benefit to be derived from them is inconsiderable; while a thorough anatomic investigation would yield far more important results. As I am working on a definite problem, I do not care at present for other forms of monsters, such as those of microcephalous, acephalous or amorphous types, but only for those which exhibit a doubling of any axial part, ranging from single individuals with doubled median parts to two complete individuals united at a single point. Similar cases