History of clinical research on the sleep apnea syndrome. The early days of polysomnography.

This article gives a brief historical overview of the literature on the sleep apnea syndrome (SAS), including initial case reports of the Pickwickian syndrome and its treatment with tracheostomy. It also highlights the beginnings of early clinical neurophysiology and describes the breakthrough in the understanding of the pathophysiology of SAS by performing polysomnography.

A long-term study on the health status and performance of sows on different feed allowances during late pregnancy. II. The total cell content and its percentage of polymorphonuclear leucocytes in pathogen-free colostrum and milk collected from clinically healthy sows.

The main objective of this study was to determine the total cell content, TCC, and the percentage of polymorphonuclear leucocytes, PMNLs, in colostrum and milk collected from sows during the first 22 days of lactation. The pH-values during the same sampling period were also determined. It should be emphasized that all the values obtained emanate from bacteriologically negative colostrum and milk. The potential influence of different levels of late gestation feeding regimes was also evaluated. The TCC-values obtained from milk samples during the first 3 weeks of lactation and exceeding the designated threshold of 10 x 10(6) cells/ml varied between 4% and 21%. Within the TCC-limitation of 10-19.99 x 10(6) cells/ml neither the preceding nor the succeeding cell counts exceeded the threshold in 26.8%. TCC-values above 19.99 x 10(6) cells/ml were preceded and succeeded by cell counts below the threshold in 58.8% and 58.8%, respectively. The TCC-levels below the threshold of 10 x 10(6) cells/ml, expressed as geometric least square means, increased significantly from day 1 to day 3 (1.23 x 10(6) cells/ml versus 1.86 x 10(6) cells/ml) and decreased thereafter gradually to day 22 (1.38 x 10(6) cells/ml). When all values were included, the TCC-values increased in a similar pattern from day 1 to day 3 (1.38 x 10(6) cells/ml versus 3.18 x 10(6) cells/ml). The value on day 22 of lactation was still on a significantly elevated level compared with that of day 1 (2.10 x 10(6) cells/ml versus 1.38 x 10(6) cells/ml). The 2 different feeding regimes were not found to influence the TCC-values during the first 22 days of lactation. In the whole material the PMNL-values, expressed as percentages of the TCC, declined from approximately 60% on day 1 of lactation to between 40% and 50% for the remaining sampling period. This decline was comparable with the one seen in the cell class below the threshold of 10 x 10(6) cells/ml. In the 2 cell classes above 9.99 x 10(6) cells/ml, 78.0% and 88.8% of PMNLs on day 1 declined to about 40% on day 22. This might indicate an inflammatory response on day 1 but without any detectable bacteriological growth. The increase in lactation number, if lactation 1 was compared with the following lactations, revealed a significant rise (p < 0.05) in TCC-level and percentage level of PMNLs. A stepwise and significant increase in pH-level occurred between days 1, 3 and 8 (6.18, 6.56, 7.03) followed by a significant decrease to day 22 (6.91) when pH-values from milk of all cell classes were included.

A long-term study on the health status and performance of sows on different feed allowances during late pregnancy. III. Escherichia coli and other bacteria, total cell content, polymorphonuclear leucocytes and pH in colostrum and milk during the first 3 weeks of lactation.

The objectives of this study were to (1) estimate the clinical status of the mammary glands and (2) compare it with the bacteriological findings, the total cell content (TCC) and its percentage of polymorphonuclear leucocytes (PMNLs) and pH in colostrum and milk secretion of sows on 2 different feeding regimes, high versus low, during late pregnancy. The milk samples were collected from both agalactia post partum (APP) sows and clinically healthy sows. Sows with a rectal temperature exceeding 39.5 degrees C within 48 h after parturition were considered to be diseased in APP and treated medically. The sows were sampled on days 1, 3, 8 and 22 of lactation during 6 consecutive lactations. Irrespective of feeding regimes, 49 out of 77 lactations among the APP sows and 15 out of 96 lactations among the clinically healthy sows revealed E. coli in pure cultures with a concomitant TCC exceeding 10 x 10(6) cell/ml already on the first day of lactation. The healthy sows with E. coli infection were denominated as being subclinically infected sows. The intensity in growth of E. coli successively declined, and the bacteria were finally eliminated between days 3 and 8 of lactation. The TCC were 82 x 10(6) cells/ml and 157 x 10(6) cells/ml in the clinically and subclinically E. coli infected glands, respectively, on the first day of sampling. The TCC declined gradually in both groups of sows, but was still higher than in bacteriologically negative milk on day 22 of lactation. The percentages of PMNLs were 66% and 79% in clinically and subclinically infected glands, respectively, on day 1 of lactation, thereafter decreasing to approximately 50% on day 22 of lactation in both groups of sows. In APP sows, swelling, reddening and/or soreness were registered in 38 out of 87 mammary glands with E. coli mastitis on the first sampling occasion. The TCC in bacteriologically negative colostrum and milk collected from APP sows on day 1 of lactation was significantly higher, 2.27 x 10(6) cells/ml, when compared with the TCC in bacteriologically negative milk secretion from the clinically healthy or subclinically infected sows, 1.38 x 10(6) cells/ml versus 1.51 x 10(6) cells/ml, respectively. The PMNLs were higher on day 1 in clinically healthy sows, 59.6%, than in subclinically infected and APP sows (43.5% and 48.3% respectively). The pH in secretion from clinically or subclinically E. coli infected glands (6.57 versus 6.46) were higher than in bacteriologically negative colostrum samples (6.29) from clinically diseased sows on the first day of sampling. On day 22 of lactation, pH-values had stabilized on a level of approximately 7.00 in all milk samples from earlier bacteriologically positive or negative mammary glands. The 2 feeding regimes, low versus high, were not found to influence TCC, PMNLs or pH except for TCC in bacteriologically negative samples of APP sows (2.69 versus 3.62). The lactation number influenced the PMNLs in both groups of sows with E. coli infected mammary glands, and both the TCC and PMNLs in bacteriologically negative colostrum and milk.

Prenatal molecular diagnosis of Gaucher disease.

Prenatal diagnosis of Gaucher disease, the most prevalent glycolipid storage disease, is based on a reliable enzyme assay of cells from amniocentesis or chorionic villous samples. However, this method cannot differentiate among the various forms of the disease. This report details four cases of prenatal diagnosis of Gaucher disease, three of which predate the use of molecular diagnosis. DNA mutation analysis to determine the genotype was predictive of the phenotypic status of the fetus and conformed to the genotype of an affected proband where available.

Failure of alglucerase infused into Gaucher disease patients to localize in marrow macrophages.

BACKGROUND: Gaucher disease is a common glycolipid storage disease, caused by a deficiency of lysosomal beta-glucosidase (glucocerebrosidase). Alglucerase is a form of glucocerebrosidase enriched with terminal mannose moieties, so as to "target" the preparation to the high-affinity macrophage receptor in patients with Gaucher disease. Our earlier in vitro studies indicated that alglucerase was bound by cells other than macrophages by a widely distributed, low-affinity mannose receptor. MATERIALS AND METHODS: Bone was removed at surgery from six patients with Gaucher disease; in three cases, bone was obtainable both when the patient was untreated and after receiving an infusion of alglucerase. Four samples of bone were obtained from patients without Gaucher disease and served as controls. A bone marrow aspirate was obtained from another patient with Gaucher disease immediately after enzyme infusion. Marrow beta-glucosidase activity and chitotriosidase (a macrophage marker) was determined on all samples. RESULTS: Even with the large bolus doses used for the treatment of Gaucher disease by some, scarcely any beta-glucosidase activity was found in marrow samples; the amount of the enzyme was much less than would have been anticipated had the enzyme been evenly distributed to all body cells. CONCLUSIONS: Alglucerase is not targeted to marrow macrophages. Its unquestioned therapeutic effectiveness must be due either to its activity at some site other than marrow macrophages or to the fact that the doses administered are so enormous that even a small fraction is sufficient to achieve a therapeutic effect.

A strategy for cloning the hereditary hemochromatosis gene.

Selective hybridization of small intestine and liver cDNA libraries was carried out using yeast artificial chromosomes (YACs) surrounding D6S105, the microsatellite that appears to be close to the gene for hereditary hemochromatosis (HFE). Of 14 candidate probes hybridizing with these YACs, only one, designated. LD5-1, detected abnormalities in southern blots of patients with hemochromatosis. Two different abnormalities. were detected in 3 of 55 patients with hemochromatosis with the LD5-1 probe, and one of these was detected in one of 44 normal subjects. The gene that hybridizes with this probe is located about 300-400 kb centromeric of D6S105. It is transcribed into mRNA that is about 8.5 kb in length in many tissues, including peripheral blood leukocytes. The available sequence indicates tha it codes for a zinc finger protein. We propose that there is a reasonable probability that LD5-1 hybridizes with the gene for hereditary hemochromatosis.

Homology between a human protein and a protein of the green garden pea.

In screening a rat mucosa cDNA subtraction library, a clone that exhibited a remarkable degree of homology with a previously described cDNA from the green garden pea, designated the 26g pea turgor protein, was found. A partial cDNA sequence from rat and a complete cDNA sequence from human were obtained. The deduced human protein had a molecular weight of 55,285 and was designated antiquitin because of its remarkable level of conservation through evolution. Human antiquitin was 60% homologous to the green pea 26g with only a single amino acid gap in each sequence. The 66 amino acids at the carboxyl ends of the human antiquitin and pea 26g proteins were 86% identical, and one segment of 52 amino acids was 92% identical. A similar partial sequence encoding 164 amino acids has been detected in Caenorhabditis elegans. Yeast DNA was found to have sequences that hybridize with a human antiquitin probe on Southern blotting. Analysis of the amount of mRNA in various rat and human tissues indicated that the largest amounts were found in rat kidney and liver and in cultured human hepatoma cells. Only minimal amounts were detected in human peripheral blood leukocytes, rat lung, or cultured human fibroblasts. Attempts to induce the mRNA by heat-shock, dehydration, ionizing irradiation, or treatment with iron, t-butylhydroperoxide, or glucocorticoids were unsuccessful. The function of the protein remains unknown.

Mutations in Jewish patients with Gaucher disease.

DNA from 100 unrelated patients, 97 of whom were Jewish and three half-Jewish, was analyzed for 22 mutations known to cause Gaucher disease. All but seven of the alleles were identified as having previously described mutations. Five of the unidentified mutations proved to be a previously undescribed nucleotide substitution in a splice junction (IVS2+1) that causes skipping of exon 2. Thus, only 2 of 197 alleles remained unidentified. Homozygotes for the most common mutation, that a nucleotide (nt) 1226, manifested, on average, the mildest disease and the latest age of onset. The mutation at nt 84 and the newly described IVS2+1 mutation, which do not produce any enzyme, were associated with earlier onset and more severe disease. Five of the mutations were considered to be "public," in the sense that they were found in more than one unrelated individual. Screening for these five mutations permitted detection of 97.5% of all Gaucher alleles in this patient population. Because the mutation at nt 1226 is underrepresented in the patient population and because not all homozygotes come to medical attention, screening the Ashkenazi population using DNA analysis should detect approximately 99% of all heterozygotes.

The origin of glucose-6-phosphate-dehydrogenase (G6PD) polymorphisms in African-Americans.

DNA samples from 54 male Afro-Americans were examined for glucose-6-phosphate dehydrogenase (G6PD) genotypes G6PD A(+)376G, G6PD A(-)202A/376G, and G6PD B and for polymorphisms in intron 5 (PvuII), at nucleotide 1311, and at nucleotide 1116 (PstI). In the G6PD B subjects, the nucleotide 1311 mutation and the PstI site appeared to be in linkage equilibrium. No PvuII+ G6PD men were encountered. The G6PD A(+) mutation was in disequilibrium with respect to both the nucleotide 1311 mutation and the PstI site. The G6PD A- nucleotide 202 mutation was in disequilibrium with all three polymorphic sites. No conclusion could be drawn with respect to the PvuII site, except that it preceded the nucleotide 202 (A-) mutation. We conclude from these and our previous studies that G6PD B is the most ancient genotype. The nucleotide 1311 mutation, with its worldwide distribution, probably occurred next. The PstI mutation, limited to Africans, probably arose next and is more ancient than the A(+) mutation, which occurred in a gene without either the PstI or the 1311 mutation. G6PD A-202A/376G is the most recent of these mutations and is still in linkage disequilibrium with all of the sites. Presumably it occurred in an individual with both the A(+) and PvuII mutations.

Glucose-6-phosphate dehydrogenase variants in Hawaii.

Sequence analysis has been performed on the DNA of 13 glucose-6-phosphate dehydrogenase (G6PD) deficient males from Hawaii, 6 of Filipino, 6 of Laotian, and 1 of Chinese extraction. Four different mutations were found: A-->T at cDNA nt 835, G-->A at nt 871, C-->T at nt 1360, and G-->A at nt 1388. The mutations at nt 835 and nt 1360 have not been described previously, and the latter, in particular, appears to be relatively common. The nt 1360 mutation changes the same codon as is altered in a previously described mutation, G6PD Andalus.

Prenatal diagnosis of glucose-6-phosphate-dehydrogenase deficiency.

Prior to the development of the DNA-based technology reliable prenatal diagnosis of G6PD deficiency was not possible. We show that, using PCR amplification and restriction endonuclease digestion, prenatal diagnosis is possible. We have now been able to determine that the male fetus of a mother heterozygous for G6PD Mediterranean had inherited the maternal X chromosome with the normal G6PD gene.

Identification of the second common Jewish Gaucher disease mutation makes possible population-based screening for the heterozygous state.

Gaucher disease is an autosomal recessive glycolipid storage disease characterized by a deficiency of glucocerebrosidase. The disease is most common in persons of Ashkenazi Jewish ancestry and the most common mutation, accounting for about 75% of the mutant alleles in this population, is known to be an A----G substitution at cDNA nucleotide (nt) 1226. Screening for this disease has not been possible because nearly 25% of the mutant alleles had not been identified, but linkage analysis led to the suggestion that most of these could be accounted for by a single mutation. We now report the discovery of this mutation. The insertion of a single nucleotide, a second guanine at cDNA nt 84 (the 84GG mutation), has been detected in the 5' coding region of the glucocerebrosidase gene. The amount of mRNA produced is shown to be normal but since the frameshift produced early termination, no translation product is seen. This finding is consistent with the virtual absence of antigen found in patients carrying this mutation. The 84GG mutation accounts for most of the previously unidentified Gaucher disease mutations in Jewish patients. The common Jewish mutation at nt 1226, the 84GG mutation, and the less-common mutation at nt 1448 accounted for 95% of all of the Gaucher disease-producing alleles in 71 Jewish patients. This now makes it possible to screen for heterozygotes on a DNA level with a relatively low risk of missing couples at risk for producing infants with Gaucher disease.

Mutation analysis of glucose-6-phosphate dehydrogenase (G6PD) variants in Costa Rica.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency has previously been reported among both the black and white populations of Costa Rica. All 28 G6PD A- samples were found to be of the common G6PD A-376G/202A type. A previously described mutation associated with nonspherocytic hemolytic anemia, G6PD Puerto Limón, was found to be due to a G----A transition at nucleotide (nt) 1192, causing a glu----lys substitution. Mutations in this region of the G6PD molecule seem invariably to be associated with chronic hemolytic anemia. G6PD Santamaria had been described previously in two unrelated white subjects. We found that both did, indeed, have the same mutations. In this variant the A----G substitution at nt 376 that is characteristic of G6PD A was present, but an A----T mutation at nt 542, apparently superimposed on the ancient G6PD A mutation, resulted in an asp----val substitution. Thus, the gain of a negative charge at amino acid 126 was counterbalanced by the loss of a charge at amino acid 181, giving rise to a variant with the G6PD A mutation but with normal electrophoretic mobility.

DNA sequence abnormalities of human glucose-6-phosphate dehydrogenase variants.

Over 400 supposedly biochemically and genetically distinct variants of glucose-6-phosphate dehydrogenase (G6PD) have been described in the past. In order to investigate these variants at the DNA sequence level we have now determined the relevant sequences of introns of G6PD and describe a method which allows us to rapidly determine the sequence of the entire coding region of G6PD. This technique was applied to six variants that cause G6PD deficiency to be functionally so severe as to result in nonspherocytic hemolytic anemia. Although the patients were all unrelated, G6PD Marion, Gastonia, and Minnesota each had identical mutations, a G----T at nucleotide (nt) 637 in exon 6 leading to a Val----Leu substitution at amino acid 213. The mutations of Nashville and Anaheim were identical to each other, viz. G----A at nt 1178 in exon 10 producing a Arg----His substitution at amino acid 393. G6PD Loma Linda had a C----A substitution at nt 1089 in exon 10, producing a Asn----Lys change at amino acid 363. The results confirm our earlier results suggesting that the NADP-binding site is in a small region of exon 10 and suggest the possibility that this area is also concerned with the binding of glucose-6-P.

Some Mexican glucose-6-phosphate dehydrogenase variants revisited.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency appears to be fairly common in Mexico. We have now examined the DNA of three previously reported electrophoretically fast Mexican G6PD variants, -G6PD Distrito Federal, G6PD Tepic, and G6PD Castilla. All three of these variants, believed on the basis of biochemical characterization and population origin to be unique, have the G----A transition at nucleotide 202 and the A----G transition at nucleotide 376, mutations that we now recognize to be characteristic of G6PD A-. Two other Mexican males with G6PD deficiency were found to have the same mutation. All five have the (NlaIII/FokI/PvuII/PstI) haplotype characteristic of G6PD A -in Africa. Since the PvuII+ genotype seems to be rare in Europe, we conclude that all of these G6PD A - genes had their ancient origin in Africa, although in many of the Mexican patients with G6PD A -202A/376G the gene may have been imported more recently from Spain, where this variant, formerly known as G6PD Betica, is also prevalent.

Definition of the mutations of G6PD Wayne, G6PD Viangchan, G6PD Jammu, and G6PD 'LeJeune'.

We report the nucleotide (nt) substitutions of four unrelated glucose-6-phosphate dehydrogenase (G6PD)-deficient males. Only the mutation of G6PD Wayne was unique. It was a nt 769 C----G substitution causing a deduced substitution of glycine for arginine at amino acid 257. This mutation is in a region in which G6PD mutations have previously been associated with chronic hemolytic anemia. The mutation of G6PD Jammu and G6PD Viangchan were identical: a G----A mutation at nucleotide 871, predicting a Val----Met substitution at amino acid 291. However, these two variants differ with respect to the 1311 polymorphism, suggesting that they may have arisen independently. Enzyme from a child with chronic hemolytic anemia, designated G6PD 'LeJeune', proved to be due to a G----T substitution at nt 637, a change identical with that in 3 unrelated patients who had been reported previously as having G6PD Gastonia, Minnesota and Marion. These findings support the suggestion that both polymorphic and sporadic G6PD deficiency mutations in unrelated persons with G6PD deficiency are often the same, even when thought to be distinct on the basis of biochemical characterization.

The NT 1311 polymorphism of G6PD: G6PD Mediterranean mutation may have originated independently in Europe and Asia.

A polymorphic site exists in exon 11 of G6PD: in the wild-type enzyme, nucleotide (NT) 1311 is a C, but is some individuals from diverse populations a T is present instead. Nine of 54 X chromosomes from Europeans of mixed origins, nine of 41 X chromosomes of Ashkenazi Jewish subjects, three of 18 X chromosomes of Sicilians, five of 20 African X chromosomes, and nine of 20 Asian Indian X chromosomes had the mutant genotype. In contrast, the mutation was found in only three of 59 Oriental X chromosomes and in three of 30 Central/South American X chromosomes. The mutation was absent from four samples of chimpanzee DNA. Twenty-one of 22 male subjects from Mediterranean countries who had the G6PD Mediterranean 563T genotype investigated in the present study or reported previously had a T at NT 1311. Only one had the normal C at NT 1311. In contrast, both G6PD Mediterranean563T males from the Indian subcontinent had the normal C at NT 1311. These findings suggest that the same mutation at nucleotide 563 giving rise to G6PD Mediterranean may have arisen independently in Europe and in Asia.

Molecular genetics of the glucose-6-phosphate dehydrogenase (G6PD) Mediterranean variant and description of a new G6PD mutant, G6PD Andalus1361A.

Glucose-6-phosphate dehydrogenase (G6PD; E.C.1.1.1.49) deficiency is the most common human enzymopathy; more than 300 different biochemical variants of the enzyme have been described. In many parts of the world the Mediterranean type of G6PD deficiency is prevalent. However, G6PD Mediterranean has come to be regarded as a generic term applied to similar G6PD mutations thought, however, to represent a somewhat heterogeneous group. A C----T mutation at nucleotide 563 of G6PD Mediterranean has been identified by Vulliamy et al., and the same mutation has been found by De Vita et al. in G6PD Mediterranean, G6PD Sassari, and G6PD Cagliari. The latter subjects had an additional mutation, at nucleotide 1311, that did not produce a coding change. We have examined genomic DNA of five patients--four of Spanish origin and one of Jewish origin--having enzymatically documented G6PD Mediterranean. All had both the mutation at nucleotide 563 and that at nucleotide 1311. A sixth sample, resembling G6PD Mediterranean kinetically but with a slightly rapid electrophoretic mobility, was designated G6PD Andalus and was found to have a different mutation, a G----A transition at nucleotide 1361, producing an arginine-to-histidine substitution. These studies suggest that G6PD Mediterranean is, after all, relatively homogeneous.