A novel mutation in the connexin 26 gene (GJB2) in a child with clinical and histological features of keratitis-ichthyosis-deafness (KID) syndrome.

BACKGROUND: Keratitis-ichthyosis-deafness (KID) syndrome is a rare congenital ectodermal disorder, caused by heterozygous missense mutation in GJB2, encoding the gap junction protein connexin 26. The commonest mutation is the p.Asp50Asn mutation, and only a few other mutations have been described to date. AIM: To report the fatal clinical course and characterize the genetic background of a premature male neonate with the clinical and histological features of KID syndrome. METHODS: Genomic DNA was extracted from peripheral blood and used for PCR amplification of the GJB2 gene. Direct sequencing was used for mutation analysis. RESULTS: The clinical features included hearing impairment, ichthyosiform erythroderma with hyperkeratotic plaques, palmoplantar keratoderma, alopecia of the scalp and eyelashes, and a thick vernix caseosa-like covering of the scalp. On histological analysis, features characteristic of KID syndrome, such as acanthosis and papillomatosis of the epidermis with basket-weave hyperkeratosis, were seen. The skin symptoms were treated successfully with acitretin 0.5 mg/kg. The boy developed intraventricular and intracerebral haemorrhage, leading to hydrocephalus. His condition was further complicated by septicaemia and meningitis caused by infection with extended-spectrum beta-lactamase-producing Klebsiella pneumoniae. Severe respiratory failure followed, and the child died at 46 weeks of gestational age (13 weeks postnatally). Sequencing of the GJB2 gene showed that the child was heterozygous for a novel nucleotide change, c.263C>T, in exon 2, leading to a substitution of alanine for valine at position 88 (p.Ala88Val). CONCLUSIONS: This study has identified a new heterozygous de novo mutation in the Cx26 gene (c.263C>T; p.Ala88Val) leading to KID syndrome.

Changes in the ultraviolet and circular dichroism spectra of aquo-, hydroxy-, azido-, and cyanocobalamin when bound to human intrinsic factor or human transcobalamin I.

The ultraviolet and the circular dichroic spectra of aquo-, hydroxy-, azido-, and cyanocobalamin free or bound to purified human intrinsic factor or purified human transcobalamin I were recorded. Except for azidocobalamin-transcobalamin I, the molar absorption for the gamma1-band in the ultraviolet spectra increased with a factor between 1.2 and 1.4 when the cobalamins were bound to protein. The ultraviolet spectrum of azidocobalamin-transcobalamin I changed so that the gamma2-band became the most intense. Minor changes were observed in the other ultraviolet spectra. The most prominent change in the CD-spectra of protein bound cobalamins was the increase in negative elipticity above 400 nm. This increase was most intense for cyano- and azidocobalamin-transcobalamin I. The study emphasizes that the mechanism by which human intrinsic factor and human transcobalamin I bind cobalamins differ.

Purification and characterization of rabbit haptocorrin.

Rabbit haptocorrin (R-binder) has been purified from serum by affinity chromatography on cobalamin-Sepharose and Blue Sepharose. It proved to be a protein with a relative molecular mass of 60 000 and an amino acid content very similar to that of other haptocorrins (Nexø, E. and Olesen, H. (1981) in B12 (Dolphin, D., ed.), Wiley Interscience, New York/London, in the press). The pH optimum (pH 6-9) for binding of cyanocobalamin and the affinity to dicyanocobinamide were like those of human and hog haptocorrins. In spectral studies, the extinction coefficient of cyanocobalamin at 363 nm (gamma 1-band) increased by about 16% on binding to rabbit haptocorrin. Binding of azidocobalamin gave spectra changes similar to those for binding to rabbit transcobalamin.

Purification and characterization of rabbit transcobalamin II.

Rabbit transcobalamin II has been purified by labile ligand affinity chromatography and G-200 Sephadex gel filtration. Structural studies indicate Stokes' radii of 2.7 nm and 3.0 nm for transocobalamin II saturated and unsaturated with cobalamin. The amino acid content of the protein is very similar to that of human transcobalamin II (Allen, R. H. (1975) Prog. Hematol. 9, 59-84). The aminoterminal sequence for transcobalamin II is reported for the first time: Glu-Ile-Cys-Gly-Val-Pro-Lys-Val-Asp-Ser-Glu-Leu-Val-Glu-Lys-Leu-Gly-Gln-Arg-Leu-Leu-Pro-(Trp)-Met-Thr). The ultraviolet and circular dichroic spectra of aquo-, hydroxo-, azido- and cyanocobalamin bound to rabbit transcobalamin II are described. On complex formation the molar absorption of the cobalamins increases and the major bands shift to longer wavelengths. The spectra are little affected by change in the fifth ligand, which indicates that the electron density around the cobalt atom is kept fairly constant by the transcobalamin II molecule. This is in contrast to the observations for the same cobalamins attached to human intrinsic factor and human transcobalamin I (Nexo, E. and Olesen, H. (1976) Biochim. Biophys. Acta 446, 143-150).

Digoxin affects potassium homeostasis during exercise in patients with heart failure.

OBJECTIVE: The aim was to evaluate whether digitalisation of heart failure patients affects extrarenal potassium handling during and following exercise, and to assess digoxin receptor occupancy in human skeletal muscle in vivo. METHODS: In a paired study of before versus after digitalisation, 10 patients with congestive heart failure underwent identical exercise sessions consisting of three bouts of increasing work rates, 41-93 W, on a cycle ergometer. The final bouts were followed by exercise to exhaustion. The femoral vessels and brachial artery were catheterised. Arterial blood pressure, heart rate, leg blood flow, cardiac output, plasma potassium, haemoglobin, pH, and skeletal muscle receptor occupancy with digoxin in biopsies were determined. RESULTS: Occupancy of skeletal muscle Na/K-ATPase with digoxin was 9% (P < 0.05). Following digitalisation femoral venous plasma potassium increased by 0.2-0.3 mmol.litre-1 (P < 0.05) at work rates of 69 W, 93 W, and at exhaustion, as well as during the first 3 min of recovery. Following digitalisation the femoral venoarterial difference in plasma potassium increased by 50-100% (P < 0.05) during exercise, and decreased by 66-75% (P < 0.05) during early recovery. Total loss of potassium from the leg increased by 138%. The effects of digitalisation on plasma potassium were not the outcome of changes in haemodynamics, because cardiac output and leg blood flow increased by up to 13% and 19% (P < 0.05), nor was it the outcome of changes in haemoconcentration or pH. CONCLUSIONS: Extrarenal potassium handling is altered as a result of digoxin treatment. This is likely to reflect a reduced capacity of skeletal muscle Na/K-ATPase for active potassium uptake because of inhibition by digoxin, adding to the reduction of skeletal muscle Na/K-ATPase concentration induced by heart failure per se. In heart failure patients, improved haemodynamics induced by digoxin may, however, increase the capacity for physical conditioning. Thus the impairment of extrarenal potassium homeostasis by heart failure and digoxin treatment may be counterbalanced by training.

Pleurodesis with polidocanol in pigs. An experimental study.

Polidocanol was instilled in five pleural cavities of three pigs. Adhesions formed in all. Adhesion distribution varied from covering a minor part to most of the lung, depending on the amount of polidocanol. One control cavity treated with sodium chloride was unaffected. Microscopy showed fibrous bridges between the pleural layers and mild submesothelial fibrosis and inflammation.

Accumulated oxygen deficit increases with inclination of uphill running.

This study examined whether accumulated oxygen deficit depends on treadmill grade during uphill running. Oxygen uptake was measured during steady-state submaximal running. By linear extrapolation at each grade, energy demand was estimated for short exhaustive runs. Oxygen deficit was the difference between this estimate and accumulated oxygen uptake. Six subjects ran at grades of 1, 15, and 20% (study I), and five males trained for anaerobic metabolism ran at 1, 10.5, and 15% (study II). Accumulated oxygen deficit was 40 +/- 11 (SD), 72 +/- 20, and 69 +/- 8 ml O2/kg, respectively (study I), and 57 +/- 8, 78 +/- 10, and 100 +/- 7 ml O2/kg (study II). The finding that accumulated oxygen deficit became larger with treadmill inclination could reflect involvement of an increasing muscle mass. However, variation in accumulated oxygen deficit was too large to make this possibility the only explanation. More likely at small treadmill inclinations energy demand for high-intensity running is underestimated by extrapolation from oxygen uptake during submaximal exercise. At high grades of uphill running, accumulated oxygen deficit reached a maximum that may reflect the subjects' anaerobic capacity for running. This hypothesis was substantiated by an enhanced accumulated oxygen deficit in the anaerobically trained subjects during 15%, but not during 1%, uphill running.

Dynamic exercise enhances regional cerebral artery mean flow velocity.

Dynamic exercise enhances regional cerebral artery mean flow velocity. J. Appl. Physiol. 78(1): 12-16, 1995.--Anterior (ACA) and middle (MCA) cerebral artery mean flow velocities (Vmean) and pulsatility indexes were determined using transcranial Doppler in 14 subjects during dynamic exercise after assessment of the carbon dioxide reactivity for both arteries. Right hand contractions provoked an elevation in left MCA Vmean [19% (12-28); P < 0.01], whereas the pulsatility decreased in all four arteries (P < 0.05). During right foot movement, left ACA Vmean increased by 23% (11-37; P < 0.01) with lesser (approximately 10%; P < 0.05) increases in the other arteries, and pulsatility index decreased (P < 0.05). During cycling, ACA and MCA Vmean increased bilaterally by 23% (10-49) and 18% (5-32), respectively (P < 0.01), and the pulsatility was also elevated (P < 0.05). Cerebral artery pulsatility did not demonstrate a focal response but depended did not demonstrate a focal response but depended on the muscle mass involved during exercise. The data demonstrate a significant increase in Vmean for the artery supplying the cortical projection of the exercising limb. Insignificant and marginally significant increases in Vmean may be related to sympathetically mediated vasoconstriction and/or coactivation of untargeted muscle groups.

Autologous fibrin sealant in reconstructive rabbit oviduct microsurgery.

A simple method for preparing concentrated fibrinogen for use in a tissue adhesive system is described. Approximately 75% to 80% of the plasma fibrinogen can be separated and concentrated within 45 to 60 minutes from a small blood sample collected from the patient before the operation. Autologous fibrinogen prepared by this method was evaluated in reconstructive microsurgery of the rabbit oviduct.

Properties and units in the clinical laboratory sciences. I. Syntax and semantic rules IUPAC--IFCC recommendations 1995.

This document is an updating of previous recommendations on the presentation of properties and their values in clinical laboratory sciences from IFCC, IUPAC and WHO. It forms part of the ongoing effort towards 'standardization' of transmission of laboratory requests and reports across cultural/language domains while avoiding standardization of the language used by clinicians or laboratory practitioners. Subsequent documents will list the kinds-of-property and the properties used in clinical laboratory sciences.

Properties and units in the clinical laboratory sciences. V. Properties and units in thrombosis and haemostasis: ISTH-IUPAC-IFCC recommendations 1995.

For historical reasons, the terms used in the nomenclature for properties in thrombosis and haemostasis differ according to 'school' of thought. This hampers communication. In collaboration, The Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis and the Committee (Commission) of Quantities and Units (in Clinical Chemistry) have prepared a set of recommended systematic names for properties in that domain. For use in electronic transmission each property has been given a code value.

International Federation of Clinical Chemistry and International Union of Pure and Applied Chemistry 2. Properties and Units in the Clinical Laboratory Sciences. VI. Properties and Units in IOC Prohibited Drugs (IFCC-IUPAC Recommendations 1997).

The term designating a substance being an active ingredient of a drug may be a generic name, a nonproprietary name, a registered trade name, a fantasy name or other. This causes difficulties in the transmission of request and report on such substances to and from the clinical laboratories, and in the collating of this information from different sources. The document comprises a list of properties of drugs of abuse in biological fluids as defined by the International Olympic Committee Medical Code for use in electronic transmission systems. Standard systematic names are presented with a code value for each. The coding schemes thus prepared are accessible on Internet from C-NPU Home page address: http://inet.uni-c.dk/ qukb7642.

International Society of Andrology and International Union of Pure and Applied Chemistry, Clinical Chemistry Section, Commission on Nomenclature, and International Federation of Clinical Chemistry Scientific Division. Properties and units in the clinical laboratory sciences XIII. Properties and units in reproduction and fertility (IUPAC-IFCC technical report 1998).

This document is the first recommendation on the presentation of properties in reproduction and fertility and their values in clinical laboratory sciences from The International Society of Andrology, IFCC and IUPAC. It forms part of the ongoing effort to standardise requests and reporting of laboratory data for transmission across cultural and linguistic domains, without attempting to standardise the language used by clinicians and laboratory practitioners. The document is accessible on Internet from C-NPU home page address: http://inet.uni-c.dk/ home/ifcc_iupac_cnpu.

Using the surgical result in the first eye to calculate intraocular lens power for the second eye.

This paper examines the possibility of using the surgical result in the first eye when planning the intraocular lens power for the second eye. Two methods were considered: (1) an empirical method by which one regards the second procedure as a repeat of the one in the first eye and calculates the power from the actual refractive error obtained in the first eye and (2) a theoretical method by which one measures the pseudophakic anterior chamber depth (ACD) of the first eye and uses this value to plan for the second eye. Based on the data from 136 second eye procedures using extracapsular cataract extraction, the prediction error of the empirical method ranged from -10.5 to +9.5 diopters. The error of the theoretical method ranged from -2.3 to +2.8 diopters, which was significantly more accurate than the empirical method (P < .001). We conclude that the fellow eye ACD may be used as a guideline for the assumed ACD of the second eye. However, such use of the fellow eye ACD could not be shown to improve power calculation predictions significantly.

Prediction of pseudophakic anterior chamber depth with the newer IOL calculation formulas.

Five methods for predicting pseudophakic anterior chamber depth (ACD) by five previously described intraocular lens power calculation formulas (Binkhorst II, Lepper and Trier, Holladay et al., Sanders-Retzlaff-Kraff (SRK/T), Olsen et al.) were evaluated in a series of 640 patients with a posterior chamber lens implant. Significant differences in formula performance were found in unusually short and long eyes. High errors were found in long eyes with the Lepper and Trier formula, the Holladay formula, and the SRK/T method. The highest accuracy was found with the Binkhorst formula and our previously described linear regression formula which expresses the pseudophakic ACD as a function of the average pseudophakic ACD for a given lens style, the preoperative ACD, and the axial length. The use of the preoperative ACD in combination with the axial length for the prediction of the pseudophakic ACD can therefore be expected to improve the accuracy of IOL power calculation.

Prediction of postoperative intraocular lens chamber depth.

The postoperative intraocular lens (IOL) chamber depth was predicted using a multiple linear regression analysis of the postoperative chamber depth as a function of the corneal height, the preoperative chamber depth, and the axial length in 279 patients with a posterior chamber lens implant. Based on a linear regression formula incorporating these preoperatively defined parameters, the postoperative IOL chamber depth could be predicted with a correlation coefficient of 0.71 and an error of +/- 0.30 mm (SD). It is concluded that an individual prediction of the IOL chamber depth will improve the accuracy in IOL calculation.

Plasma amino acid concentrations in newborn infants during parenteral nutrition.

Amino acid substance concentrations in plasma have been measured during total and partial parenteral nutrition with Vamin in 12 seriously ill newborn infants. When plasma amino acid concentrations were compared to the levels in reference infants, 11 of 21 were low, among the 11 were seven essential amino acids (Arg, Cys, Ile, Leu, Lys, Thr, Tyr), while concentrations above the reference median and range occurred for seven amino acids. For three (Asp, Glu, Phe) levels were exceptionally high. From these findings, from studies by others on amino acid levels during parenteral nutrition with Vamin, and from amino acid needs judged from venous-arterial differences in newborns, an amino acid preparation for parenteral administration to newborn infants is suggested.

Glycated hemoglobin as a measure of physiological stress.

The concentration of glycated hemoglobin (HbA1c) is an integrated expression of the blood glucose concentration level in the previous weeks. The levels of HbA1c were measured in 23 medical students who had to pass an examination within a few weeks. Twelve other students served as a control group. Four months later, the blood tests were repeated. In addition, serum cholesterol was measured. HbA1c levels were significantly higher in the exam group compared with the control group. The second test revealed a significant decrease in HbA1c in the exam group, while the values in the control group were stable. No differences in serum cholesterol were detected. It is concluded that HbA1c might be of value as a measure of physiological stress.

Human intrinsic factor. Its primary structure compared to the primary structure of rat intrinsic factor.

Human intrinsic factor was digested by trypsin and the resulting peptides purified by gelfiltration. Two peptides were sequenced to a total of 61 amino acid residues. Including the sequence for the N-terminal peptide and four cyanogen bromide peptides previously reported, we have now determined a total of 163 amino acid residues, that is a fraction of about 0.40 of the primary structure of human intrinsic factor. The 110 of the 163 residues known of human intrinsic factor are identical to the sequence of rat intrinsic factor.

Hemoglobinopathies in Danish families.

Based on cases referred for investigation, as well as a questionnaire sent to all medical and pediatric departments in Denmark, 48 cases of hemoglobinopathy in 15 families of Danish ancestry are reviewed. 18 Danes in six families have been identified as having beta-thalassemia, and remarkably one - a homozygote - has beta-thalassemia intermedia requiring treatment with iron-chelation therapy. A further 36 Danes in 9 families have a hemoglobin variant: five unstable hemoglobins (Volga, Niteroi, and three unidentified), one hereditary methemoglobinemia (M-Arhus), one polycythemia (Ty Gard) and 2 asymptomatic (Athens-Georgia and Hafnia). Although rare in Danish families, a hemoglobinopathy should be considered in families with an unexplained chronic hemolytic anemia, cyanosis or polycythemia.