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Biomedical subjects

C Holm

Publications and source records attributed to C Holm.

At least 19 recordsLinked to original sources

Tail-induced attraction between nucleosome core particles.

We study a possible electrostatic mechanism underlying the compaction of DNA inside the nuclei of eucaryotes: the tail-bridging effect between nucleosomes, the fundamental DNA packaging units of the chromatin complex. As a simple model of the nucleosome we introduce the eight-tail colloid, a charged sphere with eight oppositely charged, flexible, grafted chains that represent the terminal histone tails. We show that our complexes attract each other via the formation of chain bridges and contrast this to the effect of attraction via charge patches. We demonstrate that the attraction between eight-tail colloids can be tuned by changing the fraction of charged monomers on the tails. This suggests a physical mechanism of chromatin compaction where the degree of DNA condensation is controlled via biochemical means, namely the acetylation and deacetylation of lysines in the histone tails.

Animals↗

The hormone-sensitive lipase C-60G promoter polymorphism is associated with increased waist circumference in normal-weight subjects.

OBJECTIVE: Hormone-sensitive lipase (HSL) is a key enzyme in the mobilization of fatty acids from triglyceride stores in adipocytes. The aim of the present study was to investigate the role of the HSL gene promoter variant C-60G, a polymorphism which previously has been associated with reduced promoter activity in vitro, in obesity and type 2 diabetes. DESIGN: We genotyped two materials consisting of obese subjects and non-obese controls, one material with offspring-parents trios, where the offspring was abdominally obese and one material with trios, where the offspring had type 2 diabetes or impaired glucose homeostasis. HSL promoter containing the HSL C-60G G-allele was generated and tested against a construct with the C-allele in HeLa cells and primary rat adipocytes. HSL mRNA levels were quantified in subcutaneous and visceral fat from 33 obese subjects. RESULTS: We found that the common C-allele was associated with increased waist circumference and WHR in lean controls, but there was no difference in genotype frequency between obese and non-obese subjects. There was a significant increased transmission of C-alleles to the abdominally obese offspring but no increased transmission of C-alleles was observed to offspring with impaired glucose homeostasis. The G-allele showed reduced transcription in HeLa cells and primary rat adipocytes. HSL mRNA levels were significantly higher in subcutaneous compared to visceral fat from obese subjects. CONCLUSION: The HSL C-60G polymorphism is associated with increased waist circumference in non-obese subjects.

Adult↗

Computer simulations of the structure of colloidal ferrofluids.

The structure of a ferrofluid under the influence of an external magnetic field is expected to become anisotropic due to the alignment of the dipoles into the direction of the external field, and subsequently to the formation of particle chains due to the attractive head to tail orientations of the ferrofluid particles. Knowledge about the structure of a colloidal ferrofluid can be inferred from scattering data via the measurement of structure factors. We have used molecular-dynamics simulations to investigate the structure of both monodispersed and polydispersed ferrofluids. The results for the isotropic structure factor for monodispersed samples are similar to previous data by Camp and Patey that were obtained using an alternative Monte Carlo simulation technique, but in a different parameter region. Here we look in addition at bidispersed samples and compute the anisotropic structure factor by projecting the q vector onto the XY and XZ planes separately, when the magnetic field was applied along the z axis. We observe that the XY-plane structure factor as well as the pair distribution functions are quite different from those obtained for the XZ plane. Further, the two-dimensional structure factor patterns are investigated for both monodispersed and bidispersed samples under different conditions. In addition, we look at the scaling exponents of structure factors. Our results should be of value to interpret scattering data on ferrofluids obtained under the influence of an external field.

Journal Article↗

[Free and pedicled muscle transfer as a therapy option in urological surgery].

Various demands of reconstruction define restoration technique. The basic principle involves optimal anatomical and functional reconstruction with concurrent minimal donor-site morbidity. Dependent on the aetiology of the defect, there are various reconstructive possibilities available for obtaining an optimal result. An immediate single stage reconstruction, with the best possible result for the individual patient as the primary consideration, can be carried out by interdisciplinary teams. In this overview, the most common indications and principles of flap choice are presented in relation to the reconstructive requirements necessary for early rehabilitation of the patient or the earlier start of necessary adjuvant therapy.

Humans↗

Magnetization of polydisperse colloidal ferrofluids: effect of magnetostriction.

We exploit magnetostriction in polydisperse ferrofluids in order to generate nonlinear responses and apply a thermodynamical method to derive the desired nonlinear magnetic susceptibility. For an ideal gas, this method has been demonstrated to be in excellent agreement with a statistical method. In the presence of a sinusoidal ac magnetic field, the magnetization of the polydisperse ferrofluid contains higher-order harmonics, which can be extracted analytically by using a perturbation approach. We find that the harmonics are sensitive to the particle distribution and the degree of field-induced anisotropy of the system. In addition, we find that the magnetization is higher in the polydisperse system than in the monodisperse one, as also found by a recent Monte Carlo simulation. Thus, it seems possible to detect the size distribution in a polydisperse ferrofluid by measuring the harmonics of the magnetization under the influence of magnetostriction.

Journal Article↗

A clinical randomized study on the effects of invasive monitoring on burn shock resuscitation.

BACKGROUND: Ever since Charles Baxter's recommendations the standard regime for burn shock resuscitation remains crystalloid infusion at a rate of 4 ml/kg/% burn in the first 24h following the thermal injury. A growing number of studies on invasive monitoring in burn shock, however, have raised a debate regarding the adequacy of this regime. The purpose of this prospective, randomised study was to compare goal-directed therapy guided by invasive monitoring with standard care (Baxter formula) in patients with burn shock. PATIENTS AND METHODS: Fifty consecutive patients with burns involving more than 20% body surface area were randomly assigned to one of two treatment groups. The control group was resuscitated according to the Baxter formula (4 ml/kg BW/% BSA burn), the thermodilution (TDD) group was treated according to a volumetric preload endpoint (intrathoracic blood volume) obtained by invasive haemodynamic monitoring. RESULTS: The baseline characteristics of the two treatment groups were similar. Fluid administration in the initial 24h after burn was significantly higher in the TDD treatment group than in the control group (P = 0.0001). The results of haemodynamic monitoring showed no significant difference in preload or cardiac output parameters. Signs of significant intravasal hypovolemia as indicated by subnormal values of intrathoracic and total blood volumes were present in both treatment groups. Mortality and morbidity were independent on randomisation. CONCLUSION: Burn shock resuscitation due to the Baxter formula leads to significant hypovolemia during the first 48 h following burn. Haemodynamic monitoring results in more aggressive therapeutic strategies and is associated with a significant increase in fluid administration. Increased crystalloid infusion does not improve preload or cardiac output parameters. This may be due to the fact that a pure crystalloid resuscitation is incapable of restoring cardiac preload during the period of burn shock.

Adolescent↗

Acute hyperglycaemia following thermal injury: friend or foe?

INTRODUCTION: Hyperglycaemia and insulin resistance are common in severely burned patients, even if they have not previously had diabetes. Conventionally, hyperglycaemia is considered a part of the hypermetabolic stress response and blood glucose levels up to 215 mg/dl are tolerated before insulin therapy is initiated. Recent studies suggest that hyperglycaemia and insulin resistance are harmful and that correcting blood glucose to normal levels with insulin might improve the prognosis significantly. STUDY OBJECTIVE: The purpose of this clinical study was to evaluate blood glucose levels in severely burned patients with conventional management and to analyse the association between early hyperglycaemia and clinical outcome. DESIGN: Clinical, prospective, descriptive study. PATIENTS: Thirty seven severely burned adults (>25% total body surface area). INTERVENTIONS: Hyperglycaemia was treated according to conventional clinical practice. This included the infusion of insulin based on a blood glucose level >215 mg/dl and the maintenance of the glucose level between 180 and 200 mg/dl. MEASUREMENTS AND RESULTS: Measurements of whole-blood glucose were performed at 8, 16, 24, 36 and 48 h after the thermal injury. Additional measurements were performed if indicated. A total of 185 measurements were obtained and significant elevations of blood glucose levels (>140 mg/dl) were found in 108 (64%) of the measurements. Peak blood glucose values exceeded 140 mg/dl in all but three of the patients; however, only 17 patients received insulin treatment during the shock period. The inadequacy of the insulin treatment is shown by the mean glucose values, which exceeded 200 mg/dl in 27% of the patients. Despite a non-significant difference in the extent of burn (P=0.055), patients who died showed significantly higher maximum glucose values than patients who survived the thermal injury (P<0.05). Even though not statistically significant, blood glucose control was poorer in patients who later developed sepsis or acute renal failure (P>0.05). No correlation was found between burned surface area (TBSA) and mean plasma glucose levels during the first 48 h of resuscitation (r=0.12). CONCLUSION: Hyperglycaemia is very frequent during the resuscitation period of thermal injury and current guidelines for insulin therapy are inadequate to correct plasma glucose to normal levels. As an association between early hyperglycaemia and subsequent mortality seems to exist, more aggressive manoeuvres to reduce blood glucose may be warranted in this group of patients.

Acute Disease↗

A flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk.

AIMS: The present study describes a flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk according to the main cause of elevated counts. METHODS AND RESULTS: A total of 75 Danish bulk tank milk samples exceeding the grading level of 3.0 x 10(4) CFU ml(-1) were examined by both flow cytometry and traditional microbiological analyses. The correlation coefficient (r) between the two methods was 0.71. For the differential analyses of the dominant bacterial populations four different parameters were used to give a species-characteristic pattern. The four parameters were as follows: staining with Oregon Green conjugated wheat germ agglutinin that binds to the cell wall of bacteria, staining with hexidium iodide that binds to all bacterial DNA, the flow cytometric forward scatter and the flow cytometric side scatter. Three regions in the flow cytometric plot were defined: region 1 includes bacteria mainly associated with poor hygiene, region 2 includes psychrotrophic hygiene bacteria and region 3 includes bacteria mainly related to mastitis. The ability of the flow cytometric technique to predict the main cause of elevated bacterial counts on routine samples was examined. Comparing these results with results obtained by traditional microbiological analyses for identification showed that for 81% of the samples the two techniques agreed on the main cause of an elevated bacterial count. CONCLUSIONS: The ability of the presented flow cytometric technique to enumerate and differentiate bacteria in bulk tank milk according to the main cause of elevated counts was demonstrated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study described the first step in development of a technique suitable for routine analyses of bulk tank milk samples. A technique indicating the main cause of an elevated count will enable the farmer to eliminate the contamination source within a short time limit.

Animals↗

Predominant microflora of downgraded Danish bulk tank milk.

The microflora of downgraded Danish bulk tank milk was examined to identify the main causes of increased microbial counts. Seventy-five representative samples with a microbial count exceeding 3.0 x 10(4) cfu/mL were selected for a more detailed microbial examination. A total of 1237 isolates from these samples were identified. Gram-negative, oxidase-positive bacteria were found in 72% of the samples. Coliforms were found in 20% of the samples, and non-coliforms were found in 49% of the samples. Coryneforms, other gram-positive rods, Lactococcus spp., Micrococcus spp., and coagulase-negative Staphylococcus spp. were found in 28 to 53% of the samples. Bacillus spp., Enterococcus spp., Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus uberis, and yeasts were found in <25% of the samples. Additionally, the isolates were divided into 3 groups, based on the main cause of an elevated microbial count. Microorganisms primarily associated with poor hygiene dominated the microflora in 64% of the samples; bacteria also related to poor hygiene, but in addition associated with growth at low temperatures (psychrotrophic bacteria) dominated the microflora in 28% of the samples; and bacteria mainly associated with mastitis dominated the microflora in 8% of the samples. A bulk tank milk storage period of 48 h instead of 24 h did not affect the proportion of downgraded milk samples and could not be associated with a specific group of microorganisms. Further, no relationship was found between somatic cell counts and the presence of mastitis bacteria.

Actinomycetales↗

Molecular mechanisms regulating hormone-sensitive lipase and lipolysis.

HSL (hormone-sensitive lipase) is a key enzyme in the mobilization of fatty acids from acylglycerols in adipocytes as well as non-adipocytes. In adipocytes, catecholamines stimulate lipolysis mainly through PKA (protein kinase A)-mediated phosphorylation of HSL and perilipin, a protein coating the lipid droplet. The anti-lipolytic action of insulin is mediated mainly via lowered cAMP levels, accomplished through activation of phosphodiesterase 3B. Phosphorylation of HSL by PKA occurs at three sites, the serines 563, 659 and 660, both in vitro and in primary rat adipocytes. Phosphorylation of Ser-659 and -660 is required for in vitro activation as well as translocation from the cytosol to the lipid droplet, whereas the role of the third PKA site remains elusive. Adipocytes isolated from homozygous HSL-null mice, generated in our laboratory, exhibit completely blunted catecholamine-induced glycerol release and reduced fatty acid release, suggesting the presence of additional, although not necessarily hormone-activatable, triacylglycerol lipase(s). Basal hyperinsulinaemia, release of exaggerated amounts of insulin during glucose challenges and retarded glucose disposal during insulin tolerance tests suggest that HSL-null mice are insulin resistant. Liver, adipose tissue and skeletal muscle appear all to be sites of impaired insulin sensitivity in HSL-null mice.

Adipocytes↗

Hormone-sensitive lipase in skeletal muscle: regulatory mechanisms.

AIM: The enzymatic regulation of intramuscular triacylglycerol (TG) breakdown has until recently not been well understood. Our aim was to elucidate the role of hormone-sensitive lipase (HSL), which controls TG breakdown in adipose tissue. METHODS: Isolated rat muscle as well as exercising humans were studied. RESULTS: The presence of HSL was demonstrated in all muscle fibre types by Western blotting of muscle fibres isolated by collagenase treatment or after freeze-drying. The content of HSL varies between fibre types, being higher in oxidative than in glycolytic fibres. Analysed under conditions optimal for HSL, neutral lipase activity in muscle can be stimulated by adrenaline as well as by contractions. These increases are abolished by presence of anti-HSL antibody during analysis. Moreover, immunoprecipitation with affinity-purified anti-HSL antibody causes similar reductions in muscle HSL protein concentration and in measured neutral lipase responses to contractions. The immunoreactive HSL in muscle is stimulated by adrenaline via beta-adrenergic activation of protein kinase A (PKA). From findings in adipocytes it is likely that PKA phosphorylates HSL at residues Ser563, Ser659 and Ser660. Contraction probably also enhances muscle-HSL activity by phosphorylation, because the contraction-induced increase in HSL activity is increased by the protein phosphatase inhibitor okadaic acid and reversed by alkaline phosphatase. A novel signalling pathway in muscle by which HSL activity may be stimulated by protein kinase C (PKC) via extracellular signal regulated kinase (ERK) has been demonstrated. In contrast to previous findings in adipocytes, in muscle activation of ERK is not necessary for stimulation of HSL by adrenaline. However, contraction-induced HSL activation is mediated by PKC, at least partly via the ERK pathway. In fat cells ERK is known to phosphorylate HSL at Ser600. So, phosphorylation of different sites may explain that in muscle the effects of contractions and adrenaline on HSL activity are partially additive. In line with the view that the two stimuli act by different mechanisms, training increases the contraction-mediated, but diminishes the adrenaline mediated HSL activation in muscle. CONCLUSION: The existence and regulation of HSL in skeletal muscle indicate a role of HSL in muscle TG metabolism.

Animals↗

Monitoring free flaps using laser-induced fluorescence of indocyanine green: a preliminary experience.

In a prospective, clinical study, the clinical utility of indocyanine green for intraoperative monitoring of free tissue transfer was evaluated. The study comprised 20 surgical patients undergoing elective microsurgical procedures. Indocyanine green angiography was performed intraoperatively, immediately after flap inset, and the operating team was blind to the fluoremetric findings. Thereafter, postoperative monitoring was done exclusively by clinical examination (color, temperature, time for recapillarization, and bleeding after puncture). Final outcome was compared with results of perioperative indocyanine (ICG)-imaging, and classified either as total flap loss, partial flap loss, or successful tissue transplantation. A total of 2 (10%) complications was recorded, and included one partial and one total flap loss. Both complications were detected by intraoperative ICG imaging. Another case of intraoperative subclinical arterial spasm at the place of microvascular anastomosis was revealed by dynamic ICG-videography. This flap did not develop postoperative complications. In conclusion, evaluation of perfusion by ICG imaging is feasible in all kinds of microsurgical flaps, irrespective of the type of tissue. Even though not meeting all the criteria of an ideal monitoring device, significant additional information can be obtained. In this study, cases with arterial spasm, venous congestion, and regional hypoperfusion were revealed by intraoperative ICG-videography. There was a strong correlation between intraoperative findings and clinical outcome.

Angiography↗

Intraoperative evaluation of skin-flap viability using laser-induced fluorescence of indocyanine green.

Laser-induced fluorescence of indocyanine green (ICG) is a new method for evaluating skin perfusion, which is superior to conventional fluorescein angiography. In a prospective clinical study ICG fluorescence video-angiography was used for the intraoperative evaluation of skin-flap perfusion. The results of ICG imaging were compared with clinical outcome 1 week postoperatively. Intraoperative ICG filling defects were always associated with delayed wound healing. In 50% of the patients, the regions of sloughing and epitheliolysis corresponded accurately to the regions of dye-filling deficits. All of the flaps without ICG filling defects healed primarily. These results suggest that ICG fluorescence is a sensitive tool for assessing nutritive blood flow in pedicled skin flaps with and without an axial vessel. Future clinical studies are required to establish critical threshold fluorescence indices that correlate with skin viability in the postoperative course.

Adult↗

The effect of exercise training on hormone-sensitive lipase in rat intra-abdominal adipose tissue and muscle.

1. Adrenaline-stimulated lipolysis in adipose tissue may increase with training. The rate-limiting step in adipose tissue lipolysis is catalysed by the enzyme hormone-sensitive lipase (HSL). We studied the effect of exercise training on the activity of the total and the activated form of HSL, referred to as HSL (DG) and HSL (TG), respectively, and on the concentration of HSL protein in retroperitoneal (RE) and mesenteric (ME) adipose tissue, and in the extensor digitorum longus (EDL) and soleus muscles in rats. 2. Rats (weighing 96 +/- 1 g, mean +/- S.E.M.) were either swim trained (T, 18 weeks, n = 12) or sedentary (S, n = 12). Then RE and ME adipose tissue and the EDL and soleus muscles were incubated for 20 min with 4.4 microM adrenaline. 3. HSL enzyme activities in adipose tissue were higher in T compared with S rats. Furthermore, in RE adipose tissue, training also doubled HSL protein concentration (P < 0.05). In ME adipose tissue, the HSL protein levels did not differ significantly between T and S rats. In muscle, HSL (TG) activity as well as HSL (TG)/HSL (DG) were lower in T rats, whereas HSL (DG) activity did not differ between groups. Furthermore, HSL protein concentration in muscle did not differ between T and S rats (P > 0.05). 4. In conclusion, training increased the amount of HSL and the sensitivity of HSL to stimulation by adrenaline in intra-abdominal adipose tissue, the extent of the change differing between anatomical locations. In contrast, in skeletal muscle the amount of HSL was unchanged and its sensitivity to stimulation by adrenaline reduced after training.

Adipose Tissue↗

[Ultrasonographic scanning of patients with severe abdominal injuries].

INTRODUCTION: The aim of this study was to describe our experiences with ultrasonography (US) of severely traumatised patients, and to show whether it is a safe and fast procedure when screening for intra-abdominal lesions. MATERIAL AND METHODS: In a period from 1996 to 1999, 876 patients were received by the trauma team. The number of patients with an injury severity score (ISS) > 15, was constant during the period. Seventy-five per cent of the patients were men. One hundred and fifty-five patients (18%) died, 111 before US was carried out. RESULTS: Over the four-year period, the number of patients received by the trauma team increased from 150 in 1996 to 365 in 1999. The number of US examinations increased from 61 (1996) till 303 (1999). The number of pathological US scans was constantly about 20 per year, where as the number of pathological CT scans was between ten and 20 per year. The number of patients that went on to explorative laparotomy was between seven and 17 per year and 49 in the whole period. The sensitivity of US to detect abdominal lesions is 0.74 and the specificity 0.97, whereas that to detect free abdominal fluid is 0.88 and the specificity 0.99. DISCUSSION: US of the abdomen is a fast and safe procedure to carry out on the severely traumatised patient. An algorithm for diagnosis is suggested.

Abdominal Injuries↗

Strong electrostatic interactions in spherical colloidal systems.

We investigate spherical macroions in the strong Coulomb coupling regime within the primitive model in salt-free environment. We first show that the ground state of an isolated colloid is naturally overcharged by simple electrostatic arguments illustrated by the Gillespie rule. We furthermore demonstrate that in the strong Coulomb coupling this mechanism leads to ionized states and thus to long range attractions between like-charged spheres. We use molecular dynamics simulations to study in detail the counterion distribution for one and two highly charged colloids for the ground state as well as for finite temperatures. We compare our results in terms of a simple version of a Wigner crystal theory and find excellent qualitative and quantitative agreement.

Journal Article↗

Exon-intron organization and chromosomal localization of the mouse monoglyceride lipase gene.

Monoglyceride lipase (MGL) functions together with hormone-sensitive lipase to hydrolyze intracellular triglyceride stores of adipocytes and other cells to fatty acids and glycerol. In addition, MGL presumably complements lipoprotein lipase in completing the hydrolysis of monoglycerides resulting from degradation of lipoprotein triglycerides. Cosmid clones containing the mouse MGL gene were isolated from a genomic library using the coding region of the mouse MGL cDNA as probe. Characterization of the clones obtained revealed that the mouse gene contains the coding sequence for MGL on seven exons, including a large terminal exon of approximately 2.6 kb containing the stop codon and the complete 3' untranslated region. Two different 5' leader sequences, diverging 21 bp upstream of the predicted translation initiation codon, were isolated from a mouse adipocyte cDNA library. Western blot analysis of different mouse tissues revealed protein size heterogeneities. The amino acid sequence derived from human MGL cDNA clones showed 84% identity with mouse MGL. The mouse MGL gene was mapped to chromosome 6 in a region with known homology to human chromosome 3q21.

3' Untranslated Regions↗

A role for hormone-sensitive lipase in the selective mobilization of adipose tissue fatty acids.

The mobilization of fatty acids from rat and human fat cells is selective according to molecular structure, and notably carbon atom chain length. This study aimed at examining whether the release of individual fatty acids from triacylglycerols (TAG) by hormone-sensitive lipase (HSL) plays a role in the selectivity of fatty acid mobilization. Recombinant rat and human HSL were incubated with a lipid emulsion. The hydrolysis of 18 individual fatty acids, ranging in chain length from 12 to 24 carbon atoms and in unsaturation degree from 0 to 3 double bond(s), was measured by comparing the composition of non-esterified fatty acids (NEFA) to that of the original TAG. The relative hydrolysis (% in NEFA/% in TAG) differed between fatty acids, being about 5-fold and 3-fold higher for the most (18:1n-7) than for the least (24:0) readily released fatty acid by recombinant rat and human HSL, respectively. Relationships were found between the chain length of fatty acids and their relative hydrolysis. Among 12-24 carbon atom saturated fatty acids, the relative hydrolysis markedly decreased (by about 5- and 3-times for recombinant rat and human HSL, respectively) with increasing chain length. We conclude that fatty acids are selectively released from TAG by HSL according to carbon atom chain length. These data provide insight on the mechanism by which fatty acids are selectively mobilized from fat cells.

Adipose Tissue↗