The Causal Effect of Vitamin D Binding Protein (DBP) Levels on Calcemic and Cardiometabolic Diseases: A Mendelian Randomization Study

Background

Observational studies have shown that vitamin D binding protein (DBP) levels, a key determinant of 25-hydroxy-vitamin D (25OHD) levels, and 25OHD levels themselves both associate with risk of disease. If 25OHD levels have a causal influence on disease, and DBP lies in this causal pathway, then DBP levels should likewise be causally associated with disease. We undertook a Mendelian randomization study to determine whether DBP levels have causal effects on common calcemic and cardiometabolic disease.

Methods and Findings

We measured DBP and 25OHD levels in 2,254 individuals, followed for up to 10 y, in the Canadian Multicentre Osteoporosis Study (CaMos). Using the single nucleotide polymorphism rs2282679 as an instrumental variable, we applied Mendelian randomization methods to determine the causal effect of DBP on calcemic (osteoporosis and hyperparathyroidism) and cardiometabolic diseases (hypertension, type 2 diabetes, coronary artery disease, and stroke) and related traits, first in CaMos and then in large-scale genome-wide association study consortia. The effect allele was associated with an age- and sex-adjusted decrease in DBP level of 27.4 mg/l (95% CI 24.7, 30.0; n = 2,254). DBP had a strong observational and causal association with 25OHD levels (p = 3.2×10⁻¹⁹). While DBP levels were observationally associated with calcium and body mass index (BMI), these associations were not supported by causal analyses. Despite well-powered sample sizes from consortia, there were no associations of rs2282679 with any other traits and diseases: fasting glucose (0.00 mmol/l [95% CI −0.01, 0.01]; p = 1.00; n = 46,186); fasting insulin (0.01 pmol/l [95% CI −0.00, 0.01,]; p = 0.22; n = 46,186); BMI (0.00 kg/m² [95% CI −0.01, 0.01]; p = 0.80; n = 127,587); bone mineral density (0.01 g/cm² [95% CI −0.01, 0.03]; p = 0.36; n = 32,961); mean arterial pressure (−0.06 mm Hg [95% CI −0.19, 0.07]); p = 0.36; n = 28,775); ischemic stroke (odds ratio [OR] = 1.00 [95% CI 0.97, 1.04]; p = 0.92; n = 12,389/62,004 cases/controls); coronary artery disease (OR = 1.02 [95% CI 0.99, 1.05]; p = 0.31; n = 22,233/64,762); or type 2 diabetes (OR = 1.01 [95% CI 0.97, 1.05]; p = 0.76; n = 9,580/53,810).

Conclusions

DBP has no demonstrable causal effect on any of the diseases or traits investigated here, except 25OHD levels. It remains to be determined whether 25OHD has a causal effect on these outcomes independent of DBP. Please see later in the article for the Editors'' Summary     

Effects of body mass,temperature, and season on resting metabolism of the nocturnal gecko Hemidactylus flaviviridis

Abstract

The resting metabolic rate (RMR) of Hemidactylus flaviviridis was measured at different temperatures from 20 to 35°C during winter and summer acclimatization. The mass exponent b values ranged between 0.67 and 0.72. Winter-acclimatized geckos of various body masses had significantly lower RMRs than summer-acclimatized geckos only at 20°C. It seems that low thermal sensitivity for summer–acclimatized group may facilitate activity during its active seasons, and high thermal sensitivity between 20 and 25°C for winter–acclimatized group may conserve energy during inactivity in winter.     

The main trends in the palaeodemography of the 7th-18th century population of Latvia

The study represents palaeodemographic research of osteological material of 3304 individuals from the funds of the Anthropological Laboratory of the Institute of History of the University of Latvia in Riga, dating from the 7th to the 18th century AD. Compensated life expectancy at birth is varying between 20.3 and 22.2 years during the research period. Crude mortality has changed between 49.3 and 45% per hundred. In the early period (7th-13th century) there is a significant male prevalence (2.2-1.4); female life expectancy at the age of 20 is on average 6.6 years less than for males. This difference decreases to 5.4 years in the 13th-18th century. According to historical demography, female life span exceeded male only in the 2nd half of 19th century. The palaeodemographic data indicate that in the 7th-18th century, women in Latvia gave birth to a mean of 4-5 children (the figure includes childless women), of whom half, at most 2-2.5, reached reproductive age, on account of high child mortality. The net reproductive rate R0 (the number of descendants per individual of the parents' generation) varies between 1 and 1.25 in the study period. Concerning the completely excavated cemeteries of Lejasbiteni (7th-10h century) and Daudziesi (16th-17th century), it was possible to calculate the size and structure of the populations that had used these cemeteries. They were similar, having 45.3-49.9% of children up to an age of 14 and 24-28% individuals over the age of 30. According to historical demography, radical improvement of the demographic situation in Latvia began in the second half of the 19th century, when the process of demographic transition in Latvia started.     

Release of small transmitters through kiss-and-run fusion pores in rat pancreatic beta cells

Exocytosis of secretory vesicles begins with a fusion pore connecting the vesicle lumen to the extracellular space. This pore may then expand or it may close to recapture the vesicle intact. The contribution of the latter, termed kiss-and-run, to exocytosis of pancreatic beta cell large dense-core vesicles (LDCVs) is controversial. Examination of single vesicle fusion pores demonstrated that rat beta cell LDCVs can undergo exocytosis by rapid pore expansion, by the formation of stable pores, or via small transient kiss-and-run fusion pores. Elevation of cAMP shifted LDCV fusion pore openings to the transient mode. Under this condition, the small fusion pores were sufficient for release of ATP, stored within LDCVs together with insulin. Individual ATP release events occurred coincident with amperometric "stand alone feet" representing kiss-and-run. Therefore, the LDCV kiss-and-run fusion pores allow small transmitter release but likely retain the larger insulin peptide. This may represent a mechanism for selective intraislet signaling.     

A reexamination of the propensities of amino acids towards a particular secondary structure: classification of amino acids based on their chemical structure

The correlation between the primary and secondary structures of proteins was analysed using a large data set from the Protein Data Bank. Clear preferences of amino acids towards certain secondary structures classify amino acids into four groups: α-helix preferrers, strand preferrers, turn and bend preferrers, and His and Cys (the latter two amino acids show no clear preference for any secondary structure). Amino acids in the same group have similar structural characteristics at their Cβ and Cγ atoms that predicts their preference for a particular secondary structure. All α-helix preferrers have neither polar heteroatoms on Cβ and Cγ atoms, nor branching or aromatic group on the Cβ atom. All strand preferrers have aromatic groups or branching groups on the Cβ atom. All turn and bend preferrers have a polar heteroatom on the Cβ or Cγ atoms or do not have a Cβ atom at all. These new rules could be helpful in making predictions about non-natural amino acids.

Selective mutagenesis of lysyl residues leads to a stable and active form of delta 9 stearoyl-CoA desaturase

Stearoyl-CoA desaturase (SCD) is a short-lived integral membrane protein of the endoplasmic reticulum (ER) that catalyzes the insertion of a double bond in the delta 9 position of saturated fatty acids. Its expression has been difficult in heterologous systems. In this study, recombinant adenovirus vector was used to express both wild-type (wt) and engineered forms of rat SCD in human transformed kidney cells. In the engineered form of SCD, lysyl residues at positions 33, 35, and 36 were mutated to alanine (SCD K/A). The recombinant adenovirus also contains a cDNA encoding the green fluorescent protein (GFP). The stable reporter GFP was used to analyze the efficiency of transfection and the stability of expressed SCDs. The wt SCD was unstable upon expression, whereas expression of SCD K/A resulted in the stabilization of the protein. The proteasome inhibitor MG132 did not affect the rapid degradation of expressed wt SCD, implying that proteasome is not involved in this degradation. Functional analysis of microsomes from infected cells expressing SCD K/A resulted in the formation of holoenzyme with desaturase activity. Here we report engineering a stabilized form of a rapidly degraded membrane protein for production of an active mutant form of SCD. The adenovirus transformed cells may provide a model for the study of the effects of positive SCD expression.     

Dynamic tests and FSH biological activity in female rats with acute nephrotic syndrome

To investigate the pituitary-ovarian status during the acute state of the nephrotic syndrome, a sequence of experiments were undertaken in adult female rats after a single dose of the puromycin aminonucleoside (PAN). The functional condition of the hypophyseal-ovarian unit was determined in control and nephrotic rats by two dynamic tests. In the first one, 10 days after PAN or placebo administration female rats were stimulated with LHRH (300 ng/100 g body wt) and samples were collected at 0, 20, 40, 60 and 80 min after releasing factor administration. The second dynamic test, which was performed in control and nephrotic rats, consists of one (day 10 after PAN) or four (between days 7-10 after PAN) doses of hCG (8 UI), respectively. In all cases, serum samples were collected on day 10. In addition, the relative in vitro biological activity of FSH from control and nephrotic rats before and after LHRH stimulus was determined. The results reveal that after a stimulatory dose of LHRH the secretion of LH was significantly diminished in nephrotic rats at all registered times. By contrast, normal response was observed in terms of FSH secretion in nephrotic females. On the other hand, no ovarian response, in terms of progesterone or estradiol synthesis, was observed in nephrotic rats after either one or four stimuli with hCG Interestingly, in spite of the normal or high concentrations of FSH, the biological activity of FSH was totally abolished in nephrotic rats. On the whole, the results from this study indicate that the nephrotic syndrome had a harmful effect on the pituitary-ovarian unit, and strongly suggest that the endocrine dysfunction could be initiated at the hypophysial level; even though a specific ovarian damage is also predictable.