A case of congenital factor XIII deficiency

The case of a 53 years old woman was described in whom a congenital factor XIII deficiency was suspected because of deforming scars and hemorrhagic diathesis. A thromboelastographic declination of elasticity as well as decreased factor XIII level up to 5% of normal range were only found in all hemostatic examinations. In 2 children factor XIII decreased to half of its normal level, whereas in the youngest daughter that level was 25%. Sporadically the girl had mild diathesis. No changes in thromboelastograms were observed in members of the patient's family. The platelet function was unchanged in all examined cases.     

Smoking during pregnancy and congenital limb deficiency.

OBJECTIVE--To examine genetic and environmental factors in the origin of isolated congenital limb deficiencies. DESIGN--Case-control study with questionnaire at a family interview of cases of isolated congenital limb deficiencies (six types), negative controls (matched for age, sex, and place of residence), and positive controls (cases of sentinel anomalies). SETTING--The database of the Hungarian Congenital Abnormality Registry, 1975-84, complemented by three other sources of ascertainment (1,575,904 births). SUBJECTS--537 case-control pairs; 392 positive controls. MAIN OUTCOME MEASURES--Smoking during pregnancy, congenital limb deficiencies. RESULTS--The adjusted rate of smoking during pregnancy was significantly higher in the mothers of cases of terminal transverse defect (relative odds 1.48; 95% confidence interval 0.98 to 2.23; P = 0.017). This finding supports the hypothesis of vascular disruption as a cause of congenital limb deficiency. CONCLUSIONS--Maternal smoking during pregnancy raises the relative odds for terminal transverse limb deficiencies.     

Transfection in perfused microfluidic cell culture devices: A case study

Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.     

A case of congenital non-spherocytic hemolytic anemia caused by enzyme deficiency in pyruvate kinase

About a familial observation of PK deficiency, the authors emphasize the important clinical and biochemical heterogeneity. Interest of isotopic explorations in the therapeutic decision of splenectomy.     

A cytogenetic study on congenital limb malformations in the Japanese monkey (Macaca fuscata)

A cytogenetic investigation was performed on 88 Japanese monkeys (Macaca fuscata) with abnormal limbs from 11 free-ranging provisioned troops including nine individuals with abnormalities indistinguishable as to whether they were congenital or injurious. All of the monkeys with abnormal limbs including the nine questionable individuals had the same karyotypes as those of normal individuals. The chromosome number was 42, consisting of 20 bi-arm autosome pairs and a submetacentric X-chromosome and Y-chromosome. The ninth chromosome pair, which was the only chromosome pair with remarkable secondary constriction, displayed length polymorphism of the centromeric C-band and secondary constriction in both deformed and normal monkeys. These kinds of variants have also been commonly found in other monkey species, which have almost the same karyotype as the Japanese monkey and have not been reported to show frequent occurrence of limb malformation. We concluded therefore that chromosomal abnormalities could be excluded from the main causal factors for limb malformations of the Japanese monkey.     

Effect of transferrin on amphibian limb regeneration: a blastema cell culture study

Summary In order to study mitogenic control during axolotl limb regeneration, we have developed a primary blastema cell culture as a very sensitive bioassay for blastema mitogens. Transferrin, an iron-binding glycoprotein which has been shown to be the neurotrophic factor for muscle cells, is the mitogen which has been analysed in the present report. Addition of approximately 2 g human transferrin/ ml of serum-free culture medium enhances blastema cell proliferation 11-fold over control levels and 2-fold over that produced by the addition of nerve extracts or purified growth factors extracted from nerve tissues (basic and acidic fetal growth factor, FGF). At a higher concentration (20 g/ml), transferrin alone has no mitogenic effect unless the medium is also supplemented with FeCl₃ (100 M). The results are discussed with regard to the sensitivity of the blastema cell culture bioassay and in the context of the neurotrophic theory of urodele limb regeneration.     

Middle Eocene habitat shifts in the North American western interior: A case study

There has been great interest in the global warming events that heralded the onset of the Eocene and particularly the response of mammalian faunas to these events. However, little information is available on the subsequent deterioration of tropical habitats in the interior of North America after these major warming episodes. The decline of tropical habitats is thought to have begun during the middle Eocene in the interior of North America, but until now, no studies have been able to document the details of this event. Recent fossil collection and stratigraphic studies from sites in southwestern Wyoming and northeastern Utah that span the middle Eocene offer a unique opportunity to evaluate changes in habitat in the western interior. Using a discriminant function analysis, habitats were reconstructed for a sequence of eight stratigraphically controlled middle Eocene assemblages. Adaptive profiles (diets, substrate use, and body masses) of fossil mammal communities were statistically compared to those of extant faunas from a variety of Neotropical habitats. Previously published magnetostratigraphic data from Utah provided a means to correlate our stratigraphic sections to the geomagnetic polarity time scale and the oxygen isotope record. The discriminant model shows that there was a significant change in the mammalian community ecology near the end of the late middle Eocene that is likely reflective of a habitat shift. When correlated to the time scale and oxygen isotope record, this key transition from forested habitats typical of the tropical early Eocene to more open woodlands began about 42 million years ago in this region of the Rocky Mountains.     

A tissue culture analysis of the steps in limb chondrogenesis

Summary Tissue-culture methods can be used to test the developmental capacity of embryonic cells. In micro-mass cultures, derived from wing cells of stages 21 through 24 chick embryos, aggregates of cells form and then differentiate into cartilage nodules, as judged by the presence of an Alcian blue staining extracellular matrix. Wing cells derived from embryos as young as stage 17 can form aggregates. However, unless they are treated with db cyclic AMP and theophylline, it is not until stage 20 that these aggregates can produce cartilage in culture. In clonal cell culture, cartilage colonies are not produced by primary cell suspensions of limb cells until stage 25 when overt cartilage differentiation is occurring in vivo. It is possible to obtain clonable cartilage cells from limb cells from embryos between stages 20 and 24 if the cells are either treated with db cyclic AMP and theophylline or maintained in suspension culture for 12 to 48 hr. On the basis of these in vitro results a multiple step model for the conversion of limb mesenchyme into cartilage cells is proposed. The model involves the appearance of cells with a predisposition to form aggregates, development of the capacity to form cartilage in response to elevated levels of cyclic AMP, the appearance of receptors that translate changes in either cell shape or cell cycle parameters into elevated levels of cyclic AMP, aggregation, elevated levels of cyclic AMP, cartilage cell determination, and differentiation. This model can serve as the basis for further tests. Presented in the Opening Symposium on Nutritional Factors and Differentiation at the 28th Annual Meeting of the Tissue Culture Association, New Orleans, Louisiana, June 6–9, 1977. This work was supported by USPHS Training Grant HD00152 from the National Institute of Child Health and Human Development, while P.B.A. was a postdoctoral trainee, and by NIH Grant HD05505 to M.S.     

A case of congenital non-spherocytic hemolytic anemia caused by triose phosphate isomerase deficiency. Prenatal diagnosis

Prenatal diagnosis: The authors present a personal case of triose-phosphate-isomerase deficiency. Clinically the deficiency associates a constitutional non spherocytic anemia, paroxystic and precocius, and neuromuscular symptoms (axial hypotonia and limb palsies). A diaphragmatic paralysis may complicate the syndrome. Infections are frequent. Survival rarely goes beyond 5 years of age. Biochemical exams show the ubiquity of the deficiency. The physiopathology remains obscure. The TPI deficiency is heritable (autosomal recessive transmission). The gene has been mapped on the short arm of the chromosome 12. Prenatal diagnosis is possible.     

On the symmetry of limb deficiencies among children with multiple congenital anomalies

In humans, unpaired organs are placed in a highly ordered pattern along the left-right axis. As indicated by animal studies, a cascade of signaling molecules establish left-right asymmetry in the developing embryo. Some of the same genes are involved also in limb patterning. To provide a better insight into the connection between these processes in humans, we analysed the symmetry of limb deficiencies among infants with multiple congenital anomalies. The study was based on data collected by the International Clearinghouse for Birth Defects Monitoring Systems (ICBDMS). Registries of the ICBDMS provided information on infants who, in addition to a limb deficiency, also had at least one major congenital anomaly in other organ systems. We reviewed 815 such cases of which 149 cases (18.3 %) were syndromic and 666 (81.7 %) were nonsyndromic. The comparisons were made within the associated limb deficiencies, considering the information on symmetry, using a comparison group with malformations associated not involved in the index association. Among the non-syndromic cases, the left-right distribution of limb deficiencies did not differ appreciably between limb deficiency subtypes (e.g., preaxial, transverse, longitudinal). The left-right distribution of limb anomalies did not differ among most types of non-limb anomalies, though a predominance of left-sided limb deficiencies was observed in the presence of severe genital defects - odds ratio [OR], 2.6; 95 % CI, 1.1-6.4). Limb deficiencies (LDs) were more often unilateral than bilateral when accompanied by gastroschisis (OR, 0.1) or axial skeletal defects (OR, 0.5). On the contrary, LDs were more often bilateral than unilateral when associated with cleft lip with or without cleft palate (OR, 3.9) or micrognathia (OR, 2.6). Specifically, we found an association between bilateral preaxial deficiencies and cleft lip, bilateral amelia with gastroschisis and urinary tract anomalies, and bilateral transverse deficiencies and gastroschisis and axial skeleton defects. Of 149 syndromic cases, 62 (41.6 %) were diagnosed as trisomy 18. Out of the 30 cases of trisomy 18 with known laterality, 20 cases were bilateral. In the remainder the right and left sides were equally affected. Also, in most cases (74.4 %) only the upper limbs were involved. In conclusion the left-right distribution of limb deficiencies among some non-limb anomalies may suggest a relationship between the development of the limb and the left-right axis of the embryo.     

Interaction of cell culture with downstream purification: a case study

Separation of product from secreting mammalian cells in the culture broth means the transition from product generation to product isolation. This interface within a biotech production process has to perform a proper solid/liquid phase separation of the cell suspension to make the product containing fluid amenable for further purification. These subsequent steps require fluid with low occurence of contaminants in order to function properly. The goal of this study was to evaluate some economic and fast cell separation methods for the preparation of a product fluid ready for use in further ultrafiltration and chromatographic processes. We have performed experiments to test the usefulness of disc stack centrifuges and tangential flow microfiltration units at large scale. Both systems revealed outstanding prospects with regard to throughput and scale up properties. However, the centrificgation did not lead to a fluid sufficiently free of particles for direct ultrafiltration or chromatography. Thus, an additional filtration step was necessary. On the other hand microfiltration led to an acceptable quality of process fluid directly. By optimisation of process parameters an effective, reproducible and robust cell separation can be obtained. However, our experience has been that such optimal conditions are somewhat specific for a narrow range. Thus, even the equipment functioning well with one type of cell would possibly not perform as well with another cell or even with the same cell under conditions slightly different to the usual situation.     

Etiological aspects of congenital diaphragmatic hernia: results of a case comparison study

We report the results of a parental questionnaire concerning possible etiological and teratological factors, such as exposure to herbicides, in the development of congenital diaphragmatic hernia (CDH). The herbicide Nitrofen interferes with lung development in rats, can induce diaphragmatic hernia and greatly resembles thyroid hormone. No association with the studied teratogens nor with maternal thyroid dysfunction was found. The questionnaire was completed by 33 parents whose baby had CDH, and by 43 couples whose baby had oesophageal atresia. The resemblance of Nitrofen to thyroid hormone, a well-known growth factor for the developing lung, is of particular interest from a pathogenetic point of view in the development of CDH.     

Yellow leaf spot of cashew: A case of molybdenum deficiency

Summary Rooted cuttings ofSeverinia buxifolia were inoculated with the vesicular-arbuscular mycorrhizal (VAM) fungusGlomus intraradices or provided an inoculum filtrate (non-VAM plants) and grown in one of seven media combinations of fired montmorillonite clay (FMC) and Canadian peatmoss (CP) at ratios of 100%, 80%, 67%, 50%, 33%, 20%, or 0% FMC. Mycorrhizal infection increased with higher proportions of FMC, but the growth of both VAM and non-VAM plants was reduced with increased FMC amendment. The growth benefit (top and root fresh-dry weights) conferred by mycorrhizal infection was greater at higher levels of FMC in the media. Improved phosphorus uptake by inoculated severinia plants appeared at least partially responsible for increased growth compared to non-VAM plants under conditions of high soluble salts and pH associated with high FMC composition. Florida Agr. Expt. Sta. Journal Series No. 6319.     

A clinical and experimental overview of sirenomelia: insight into the mechanisms of congenital limb malformations

Sirenomelia, also known as sirenomelia sequence, is a severe malformation of the lower body characterized by fusion of the legs and a variable combination of visceral abnormalities. The causes of this malformation remain unknown, although the discovery that it can have a genetic basis in mice represents an important step towards the understanding of its pathogenesis. Sirenomelia occurs in mice lacking Cyp26a1, an enzyme that degrades retinoic acid (RA), and in mice that develop with reduced bone morphogenetic protein (Bmp) signaling in the caudal embryonic region. The phenotypes of these mutant mice suggest that sirenomelia in humans is associated with an excess of RA signaling and a deficit in Bmp signaling in the caudal body. Clinical studies of sirenomelia have given rise to two main pathogenic hypotheses. The first hypothesis, based on the aberrant abdominal and umbilical vascular pattern of affected individuals, postulates a primary vascular defect that leaves the caudal part of the embryo hypoperfused. The second hypothesis, based on the overall malformation of the caudal body, postulates a primary defect in the generation of the mesoderm. This review gathers experimental and clinical information on sirenomelia together with the necessary background to understand how deviations from normal development of the caudal part of the embryo might lead to this multisystemic malformation.