LDL cholesterol level as a risk factor for retinopathy and nephropathy in children and adults with type 1 diabetes mellitus: A nationwide cohort study.

BACKGROUND: Microvascular complications are common in people with diabetes, where poor glycaemic control is the major contributor. The aim of this study was to explore the association between elevated LDL cholesterol levels and the risk of retinopathy or nephropathy in young individuals with type 1 diabetes. METHODS: This was a nationwide observational population-based cohort study, including all children and adults with a duration of type 1 diabetes of ≤ 10 years, identified in the Swedish National Diabetes Register between 1998 and 2017. We calculated the crude incidence rates with 95% confidence intervals (CIs) and used multivariable Cox regression to estimate crude and adjusted hazard ratios (HRs) of retinopathy or nephropathy in four LDL cholesterol categories: <2.6 (Reference), 2.6-3.4, 3.4-4.1 and > 4.1 mmol L-1 . RESULTS: In total, 11 024/12 350 (retinopathy/nephropathy, both cohorts, respectively) children and adults (median age 21 years, female 42%) were followed up to 28 years from diagnosis until end of study. Median duration of diabetes when entering the study was 6 and 7 years in the retinopathy and nephropathy cohort, respectively. Median LDL cholesterol was 2.4 mmol L-1 , and median HbA1c level was 61 mmol mol-1 (7.7 %). After multivariable adjustment, the HRs (95% CI) for retinopathy in individuals with LDL cholesterol levels of 2.6-3.4, 3.4-4.1 or > 4.1 mmol L-1 were as follows: 1.13 (1.03-1.23), 1.16 (1.02-1.32) and 1.18 (0.99-1.41), compared with the reference. The corresponding numbers for nephropathy were as follows: 1.15 (0.96-1.32), 1.30 (1.03-1.65) and 1.41 (1.06-1.89). CONCLUSIONS: Young individuals with type 1 diabetes exposed to high LDL cholesterol levels have an increased risk of retinopathy and nephropathy independent of glycaemia and other identified risk factors for vascular complications.

Effect of hatching time on time to first feed intake, organ development, enzymatic activity and growth in broiler chicks hatched on-farm.

The conventional commercial hatcheries used today do not allow the newly hatched chicks to consume feed or water. Combined with natural variation in hatching time, this can lead to early hatched chicks being feed-deprived for up to 72 h before being unloaded at the rearing site. This study investigated the effects of hatching time on time to first feed intake and development of organs, digestive enzymes and productivity in terms of growth and feed conversion ratio in chicks hatched on-farm. Chicks were divided into three hatching groups (early, mid-term and late), and assessed over a full production cycle of 34 days. The results revealed that chicks remain inactive for a considerable amount of time before engaging in eating-related activities. Eating activity of 5% (i.e. when 5% of birds in each hatching group were eating or standing close to the feeder) was recorded at an average biological age (BA) of 25.4 h and a proportion of 50% birds with full crop was reached at an average BA of 30.6 h. Considering that the hatching window was 35 h in this study, the average chick probably did not benefit from access to feed and water immediately post-hatch in this case. At hatch, mid-term hatchlings had a heavier small intestine (30.1 g/kg bw) than both early (26.4 g/kg bw) and late (26.0 g/kg bw) hatchlings. Relative length of the small intestine was shorter in late hatchlings (735 cm/kg bw) than in mid-term (849 cm/kg bw) and early (831 cm/kg bw) hatchlings. However, the relative weight of the bursa fabricii was greater in mid-term (1.30 g/kg bw) than in early hatchlings (1.01 g/kg bw). At hatch, late hatchlings were heavier than early and mid-term hatchlings (P < 0.05), but by 3 days of age early hatchlings were heavier than mid-term and late hatchlings (P < 0.01). The only effect persisting throughout the study was a difference in the relative weight of the small intestine, where late hatchlings had heavier intestines than early hatchlings (P < 0.05). Thus, while there were differences between hatching groups, this study showed that the hatchlings seemed capable of compensating for these as they grew.

Comparison of photocatalysis and photolysis of malathion, isomalathion, malaoxon, and commercial malathion--products and toxicity studies.

Malathion, one of the most widely applied insecticides, is still used in agriculture. There are many studies regarding its degradation under different experimental conditions, but few deal with its transformation products, i.e. malaoxon and isomalathion. Thus, malathion, malaoxon, isomalathion, and Radotion (one of its over 6000 commercial forms) were studied in terms of their degradation kinetics, identification of their transformation products, their toxicity, and their degree of mineralization, during UV photolysis (lambda = 254 nm) and TiO(2) photocatalysis (lambda = 355 nm). The degradation kinetics was similar for all four starting materials. More than 75% of theoretically expected sulfur in PS and P-S groups was oxidized after 240 min of photolysis and photocatalysis. On the other hand, less than 30% of stoichiometrically predicted amounts of phosphate was detected in the photolytic experiments, but more than 80% of expected phosphate was detected after photocatalytic treatment of all four organophosphorous materials. Several transformation products were identified by mass spectra of representative gas chromatographic peaks. Oxidation and isomerization were found as the main reactions of butenedioc acid diethyl esters and their analogs. The formation of malaoxon, isomalathion or trimethyl phosphate esters correlated well with the induced toxicity (inhibition of acetylcholinesterase), which was observed in photocatalysis of malathion and Radotion, and in photolysis of malaoxon and Radotion.

Photodegradation of organophosphorus insecticides - investigations of products and their toxicity using gas chromatography-mass spectrometry and AChE-thermal lens spectrometric bioassay.

Four organophosphorus compounds: azinphos-methyl, chlorpyrifos, malathion and malaoxon in aqueous solution were degraded by using a 125 W xenon parabolic lamp. Gas chromatography-mass spectrometry (GC-MS) was used to monitor the disappearance of starting compounds and formation of degradation products as a function of time. AChE-thermal lens spectrometric bioassay was employed to assess the toxicity of photoproducts. The photodegradation kinetics can be described by a first-order degradation curve C=C0e(-kt), resulting in the following half lives: 2.5min for azinphos-methyl, 11.6 min for malathion, 13.3 min for chlorpyrifos and 45.5 min for malaoxon, under given experimental conditions. During the photoprocess several intermediates were identified by GC-MS suggesting the pathway of OP degradation. The oxidation of chlorpyrifos results in the formation of chlorpyrifos-oxon as the main identified photoproduct. In case of malathion and azinphos-methyl the corresponding oxon analogues were not detected. The formation of diethyl (dimethoxy-phosphoryl) succinate in traces was observed during photodegradation of malaoxon and malathion. Several other photoproducts including trimethyl phosphate esters, which are known to be AChE inhibitors and 1,2,3-benzotriazin-4(3H)-one as a member of triazine compounds were identified in photodegraded samples of malathion, malaoxon, and azinphos-methyl. Based on this, two main degradation pathways can be proposed, both result of the (P-S-C) bond cleavage taking place at the side of leaving group. The enhanced inhibition of AChE observed with the TLS bioassay during the initial 30 min of photodegradation in case of all four OPs, confirmed the formation of toxic intermediates. With the continuation of irradiation, the AChE inhibition decreased, indicating that the formed toxic compounds were further degraded to AChE non-inhibiting products. The presented results demonstrate the importance of toxicity monitoring during the degradation of OPs in processes of waste water remediation, before releasing it into the environment.

The association between the serum concentration of canine prostate specific esterase (CPSE) and the size of the canine prostate.

Benign prostatic hyperplasia (BPH) is an age-related disorder in the intact male dog that is associated with an increase in the prostatic size. Ultrasonography gives a reliable estimate of the prostatic size, but a method for screening the prostate size using a serum sample has advantages, such as requiring less expensive equipment. The primary aim of the study was to study the association between the concentration of the circulating biomarker canine prostate specific esterase (CPSE) and prostatic size. Seventy-nine dogs that were four years old or older were included in the study. Ultrasonography was used for calculating the volume of the prostate. The calculated volume was divided by an estimate of the normal prostatic volume in dogs aged one to four years, to determine the relative prostatic size: the size of the prostate in relation to the normal size in dogs 1-4 years old (Srel). CPSE was analyzed from serum samples. Multiple linear regression analysis was used for studying associations between variables. Prediction intervals for the relative prostatic size based on CPSE concentrations were calculated, as were receiver operating curves for CPSE concentrations predicting Srel. The concentration of CPSE was associated with the relative size and contour of the prostate (P < 0.001). All dogs with clinical signs of BPH had an Srel ≥ 2.5. A CPSE concentration of 200 ng/mL predicted Srel to 2.5 (95% P.I: 1.2-4.8). Based on ROC analysis, the optimal discrimination threshold for CPSE concentration for Srel ≥ 2.5 was estimated as 90 ng/mL (95% confidence interval: 50-140), with a sensitivity of 85% and a specificity of 72%. Screening for CPSE is of potential value in the aging intact male dogs. Although many dogs with an Srel ≥ 2.5 show no clinical signs, the insidious nature of BPH supports further investigations of the prostate in these dogs, corresponding to a CPSE concentration of approximately 90 ng/mL or higher.

The sequential development of spongiform change and gliosis of scrapie in the golden Syrian hamster.

The lesion profiles of spongiform change and gliosis in the hamster occurring after intracerebral (IC) inoculation of scrapie virus, are calculated and compared to the lesion profile of spongiform change of scrapie in mice and of scrapie and Creutzfeldt-Jakob disease (CJD) in the squirrel monkey. The profile of scrapie in hamsters differs considerably from that of a closely related strain of scrapie in mice, and both differ from scrapie and CJD in the squirrel monkey. These differences emphasize the effect of the host on the distribution of pathological changes in these unconventional virus infections. The sequential development of the lesions in the hamster shows that the earliest changes are detectable before the onset of clinical disease 49-57 days after inoculation, as assessed by light microscopy. Gliosis is detectable by indirect immunofluorescence 35-39 days after inoculation by use of a monoclonal antibody directed against astrocytes.

Monoclonal antibodies to central nervous system antigens.

Thirty monoclonal antibodies produced by mouse hybrid myelomas which react with antigens in hamster or mouse nervous system tissues were derived. Using these antibodies as probes with indirect immunofluorescence and immunoperoxidase techniques, we can selectively identify by morphological criteria many of the structural components of the brain seen at a light-microscopic level, including the neutrophil, neuronal cytoplasm, nuclei, axons, astrocytes and ependyma. Some of the antibodies display cytoskeletal and filamentous structures, including intermediate filaments, microfilaments, neurofilaments, glial and ependymal filaments. The specificity to neural tissue components of these hybridoma antibodies was assessed by their reactivity to mouse and hamster non-neural tissues and selected mouse, hamster, rat and human cultured cell lines. Of the 30 clones analyzed, specificity ranged from 3 clones reacting only with grey matter of mouse and hamster brain, one clone reacting only with axons in animal and human brain, to 19 clones reactive with both neural and non-neural tissue components.

Site-directed antisense oligonucleotide decreases the expression of amyloid precursor protein and reverses deficits in learning and memory in aged SAMP8 mice.

beta amyloid protein (Abeta) is a 40-43 amino acid peptide derived from amyloid precursor protein (APP). Abeta has been implicated as a cause of Alzheimer's disease (AD). Mice with spontaneous or transgenic overexpression of APP show the histologic hallmarks of AD and have impairments in learning and memory. We tested whether antisense phosphorothiolated oligonucleotides (AO) directed at the Abeta region of the APP gene given with or without antibody directed at Abeta could reverse the elevated protein levels of APP and the behavioral impairments seen in SAMP8 mice, a strain which spontaneously overexpresses APP. We found that intracerebroventricular (ICV) administration of antibody with either of two AOs directed at the midregion of Abeta improved acquisition and retention in a footshock avoidance paradigm, whereas two AOs directed more toward the C-terminal, a random AO, and vehicle were without effect. Three injections of the more potent AO given without antibody reduced APP protein levels by 43-68% in the amygdala, septum, and hippocampus. These results show that AO directed at the Abeta region of APP can reduce APP levels in the brain and reverse deficits in learning and memory.

Determination of organophosphate and carbamate pesticides in spiked samples of tap water and fruit juices by a biosensor with photothermal detection.

The determination of organophosphate (paraoxon, chlorpyrifos, diazinon) and carbamate (carbaryl, carbofuran) pesticides in spiked drinking water and fruit juices was carried out using a photothermal biosensor. The biosensor consists of a cartridge containing immobilised enzyme acetylcholinesterase (AChE) placed in a flow-injection analysis (FIA) manifold and a photothermal detector based on thermal lens spectrometry. With this approach, 0.2 ng/ml of paraoxon can be detected in less than 15 min. Limits of detection for other organophosphate (chlorpyrifos, diazinon) and carbamate (carbaryl, carbofuran) pesticides varied, depending on their antiacetylcholinesterase (AntiAChE) toxicity, from 1 ng/ml to 4 microg/ml. The biosensor was used for the direct detection of pesticides in spiked tap water and fruit juices without any pretreatment steps. In these cases, the LOD3sigma of 1.5, 2.8 and 4 ng/ml paraoxon in tap water, orange juice and apple juice were obtained, respectively.

Loop arrays in mouse brain demonstrated with antisera to cytokeratins and monoclonal antibodies to several classes of intermediate filaments: strain differences and developmental expression.

Some monoclonal antibodies raised against mouse brain antigens display a novel loop array apparently localized within the cytoplasm of neurons in fresh frozen sections of adult mouse brain. By indirect immunofluorescence, these loops are detectable in the cerebral cortex, thalamus, brainstem, and are particularly striking in association with pyramidal neurons of the hippocampus. The loops are also seen with polyclonal antibodies to the cytokeratin class of intermediate filaments. The antibodies which react with these loops also react with ependymal cells. Western blot analysis of crude insoluble cytoskeletal components of mouse brain with antibodies of cytokeratins confirm the presence of reactive bands in the range of 40-60 kdalton, appropriate in molecular weight for this class of cytoskeletal filaments. This evidence suggests that the loops share antigenic determinants with non-neural cytokeratins. During development, immunoreactive structures are first seen as small punctate or curvilinear profiles, which change into a loop array at approximately 14 days postnatal age in several mouse strains. However, in 8 of 15 different mouse strains, these immature punctate profiles remain without morphological alteration to loops throughout adult age. The F1 crosses between strains with and without the loops develop loops, but on average they are of smaller size than in the positive parent.

Effect of metformin on nitric oxide synthase in genetically obese (ob/ob) mice.

Genetically obese (ob/ob) mice were employed for the study of the effect of metformin on activity and expression of nitric oxide synthase (NOS ) in vitro and in vivo. For in vitro analysis, mouse liver extracts were used. For the in vivo study, (ob/ob) and their control litter mates (ob/c) mice were injected with specified amounts of metformin and the expression of NOS in the adipose tissue and hypothalamus was measured by Western blotting. Results show that metformin exhibited a biphasic effect on NOS activity in vitro. Expression of metformin was differentially altered in the hypothalamus and adipose tissues of the normal and ob/ob animals that were treated with metformin. Further, a significant decrease in food intake occurred in the (ob/ob) mice that received metformin. This decrease in food intake was not accompanied by changes in serum glucose. At inhibitory concentrations, hypothalamic NOS expression changes differentially in normal and ob/ob mice. In normal mice, metformin stimulated NOS expression, while in ob/ob mice there was an inhibition. NOS expression increased in brown adipose tissue of metformin treated control mice, while no such increase was observed in ob/ob mice. No effect of metformin was observed in white adipose tissue of control or obese mice. Thus, metformin may produce anorectic effects through modulation of NOS.

Effect of organic solvents in the on-line thermal lens spectrometric detection of chromium(III) and chromium(VI) after ion chromatographic separation.

The effect of methanol, acetone and acetonitrile on the sensitivity, selectivity and the detection limits (LODs) of the determination of chromium species by ion chromatography was investigated. A collinear dual-beam thermal lens spectrometer was used for the direct detection of chromium complexes [pre-column derivatized Cr(III)-pyridine-2,6-dicarboxylic acid, and post-column derivatized Cr(VI)-1,5-diphenylcarbazide] following the ion chromatographic separation on a Dionex HPIC-CS5A solvent compatible column. Different amounts of organic solvents were added directly to the eluent (up to 30%) and to the post-column reagent (up to 60%) to improve the thermooptical properties of the solvents. Consequently, the sensitivity of the technique was increased by a factor of 2-3 and LODs of 0.1 and 10 microg dm(-3) were achieved for Cr(VI) and Cr(III), respectively, when the eluent reaching the detector contained 30% of acetonitrile. The addition of organic solvents also resulted in significant changes in retention times, which improved the Cr(III)/Cr(VI) separation.

Ultrasensitive determination of beta-carotene in fish oil-based supplementary drugs by HPLC-TLS.

This research study demonstrates that thermal lens spectrometry (TLS) combined with efficient isocratic HPLC separation provides an excellent analytical means to profile and quantify carotenes in fish body oils and possibly also in vegetable oils. In particular, a highly sensitive, selective, simple and rapid method for the determination of ultratraces of beta-carotene in fish oil-based supplementary drugs has been proposed. The analyte could be determined reliably and precisely in the presence of a complex matrix including other fat-soluble vitamins (A, D and E), polyunsaturated fatty acids, sterols and other pigments by matrix-matched calibration in cod liver oil. The suitability of the method is also evidenced by favourable analytical figures of merit for beta-carotene such as a linearity range of 1-120 ng ml(-1), LOD of 0.58 ng ml(-1), recovery of 101.4+/-3.3% and measurement repeatability of 4.1%.

Days-lost to training and competition in relation to workload in 263 elite show-jumping horses in four European countries.

Orthopaedic, or other, injuries in sports medicine can be quantified using the 'days-lost to training' concept. Both the training regimen and the surface used in training and racing can affect the health of racehorses. Our aim was to associate 'days-lost to training' in elite-level show-jumpers to horse characteristics, training and management strategies, and the time spent working on various training and competition surfaces. We designed a longitudinal study of professional riders in four European countries. Data were recorded using training diaries. Reasons for days-lost were classified into non-acute and acute orthopaedic, medical, hoof-related, and undefined. We produced descriptive statistics of training durations, relative to type of training, surfaces used, and days-lost. We created zero-inflated negative-binomial random-effects models using the overall days-lost as outcome. In the whole dataset, duration variables related to training surfaces were analysed as independent. The Swedish data only were also used to test whether duration variables were related to competition surfaces. Thirty-one riders with 263 horses provided data on 39,028 days at risk. Of these, 2357 (6.0%) were days-lost (55% and 22% of these were due to non-acute and acute orthopaedic injuries, respectively) in 126 horses. In the all-country model, controlling for season, a significant variable was country. Switzerland and the UK had lower incidence-rate ratios (IR) compared to Sweden (IRs 0.2 and 0.03, respectively). Horses with previous orthopaedic problems had almost a doubled IR (1.8) of days-lost due to orthopaedic injury, compared to baseline. If the horse had jumping training more than 1 min per day at risk the IRs were 6.9-7 (compared to less than this amount of time); this was, however, likely an effect of a small baseline. Variation in training was a protective factor with a dose-response relationship; the category with the highest variation had an IR of 0.1. In the Swedish model, controlling for season, there was an association of year (IR 2.8 year 2010). Further, if the horse rested >17-25% of the days at risk, or >33% of the DAR2, had IRs 3.5 and 3.0, compared to less time. Horses ≥ 6 years had IRs of 1.8-2.0, compared to younger horses. Limited training use of sand surface was a risk-factor (IR 2.2; >4 ≤ 12 min/day at risk), compared to not training on sand. Training/competing on sand-wood was a protective factor (IRs 0.4-0.5) compared to not using this surface.

Replantation of a vertically fractured maxillary central incisor after repair with adhesive resin.

AIM: To describe the replantation of a vertically fractured root-filled maxillary central incisor after repair with adhesive resin. SUMMARY: Root-filled teeth with vertical root fractures are usually extracted as they generally have a poor prognosis. In this case, an 18-month follow-up of a replanted incisor with vertical root fracture is presented with clinical and radiographic documentation. KEY LEARNING POINTS: * Replantation of root-filled incisors with complete vertical root fracture after resin bonding might be a realistic alternative to extraction. * Practitioners should be aware of the possible disadvantages as well as advantages associated with failure of a replanted incisor with complete vertical root fracture.

Recent applications of thermal lens spectrometry in food analysis and environmental research.

This review summarizes the most recent achievements related to the application of thermal lens spectrometry (TLS) in food analysis and environmental research. All the applications are associated with the use of an appropriate analytical procedure providing sufficient selectivity, that cannot be achieved by TLS itself. Several selective reagents, biosensors and chromatographic separation procedures (IC, HPLC), used for this purpose, and their performance in combination with TLS, are described. Heavy metals and related species, pesticides, carotenoids, fatty acids, and their determination in samples such as water, fruit juices, oils and marine phytoplankton were given most consideration. The main advantages of novel analytical methods include improved sensitivity and selectivity, simplicity, minimized need for sample preparation and handling as well as reduced time of analysis.

Experimental considerations of simultaneous thermal lens and beam deflection phenomena.

A theoretical model, believed to be novel, that describes simultaneous thermal lens and beam deflection phenomena was experimentally tested. The effect of beam deflection on thermal lens measurements and the effect of the thermal lens on beam deflection measurements were investigated. The experiment confirmed the validity of theoretical predictions for measurements of strongly absorbing samples for which the assumption of an unchanged pump beam in the sample is still valid.

Optimisation of FIA system for detection of organophosphorus and carbamate pesticides based on cholinesterase inhibition.

The sensitivity of the bioanalytical FIA system containing different immobilised cholinesterases (AChEs from electric eel, human erythrocytes, bovine erythrocytes and BuChE from horse serum) for determination of organophosphorus and carbamate pesticides was tested. Responses to some frequently used organophosphorus (paraoxon, oxydemeton-methyl, triazophos) and carbamate (carbofuran, propoxur) pesticides were found to be dependent on the origin of cholinesterases. The highest sensitivity was obtained by bioanalytical columns prepared with electric eel AChE while the lowest sensitivity was shown by the bioanalytical columns prepared with horse serum BuChE. The differences in responses for different enzymes were found to be less pronounced when the contact time between the enzyme and the pesticide is long enough (low flow rates). The optimal flow rate was chosen as a compromise between the duration of analysis and reasonably low limits of detection.

Determination of trans-beta-carotene and other carotenoids in blood plasma using high-performance liquid chromatography and thermal lens detection.

Thermal lens spectrometry (TLS) was applied for the detection of beta-cryptoxanthin, alpha-carotene, trans-beta-carotene, and lycopen in blood plasma. This combined high-performance liquid chromatography-TLS (HPLC-TLS) method was validated by comparison with HPLC-UV-Vis analysis of blood plasma under identical chromatographic conditions and by comparing the results obtained from an independent, standard HPLC procedure for determination of carotenoids in blood plasma samples. The results demonstrated good agreement with the target values for carotenoids in an in-laboratory control sample and confirmed the accuracy of the HPLC-TLS technique. Limits of detection for blood plasma samples were 70 pg/ml for beta-cryptoxanthin, 85 pg/ml for alpha-carotene, 100 pg/ml for trans-beta-carotene, and 120 pg/ml for lycopen. This represents a 100-fold improvement compared to the HPLC analysis with UV-Vis detection.