In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network.

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time-period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post-thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital® (SV) technology, the first commercialized production line of SpermVital® (C) and by conventional procedure applying Biladyl® extender (B). Post-thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital® technology is improved and is more efficient in conserving post-thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.

Reproductive performance of immobilized cryopreserved bovine semen used for timed artificial insemination.

The SpermVital® technology comprises embedding of spermatozoa within an alginate gel to facilitate release of sperm cells over a prolonged period in utero after AI. The aim of this study was to examine whether the survival time of spermatozoa is extended when applying this immobilization technology in combination with cryopreservation. Sperm cell survival (acrosome and plasma membrane integrity) was studied in vitro for 48 hr at physiological temperature. One dose of SpermVital® (SV) semen was compared with single doses of Biladyl® (B) processed semen as well as double doses of B (B double). B double was obtained by adding a second B dose the following day, thereby mimicking double AI. Furthermore, reproductive performance applying single early timed AI (TAI) with SV following oestrus synchronization was studied in a field trial. Double insemination (TAI on two consecutive days) with B semen served as control. Number of acrosome-intact live sperm cells decreased over time in vitro for all treatments (p < .05). There was no difference between SV sperm cell survival and B double after 24 hr (p > .05). However, after 48 hr, SV sperm cell survival was higher than B double (p < .05). Moreover, multivariate analysis showed that the outcome of single early TAI with SV was not significantly different from B double (p > .05). Likelihood of pregnancy and calving in the heifer group was higher than in the cow group (p < .05). These results imply that spermatozoa immobilized in alginate gel have prolonged survival.

Effect of changes in an FFQ: comparing data from two national dietary survey instruments among 2-year-olds.

In the interpretation of dietary trends, it is important to consider the potential effect of modifications in the dietary assessment method. Therefore, our objective was to explore the comparability of data obtained at two time points by a semi-quantitative FFQ (SFFQ) which has had slight modifications over time. In the national dietary surveys among Norwegian 2-year-olds, diet was assessed by an SFFQ which underwent modifications between the 1999 survey and the 2007 survey. In the present study, fifty-nine families with a 2-year-old child participated by completing both the SFFQ in a crossover design within a month's time. With regard to the reported intake of energy and nutrients, the largest significant differences observed between the two questionnaires were for carbohydrates and added sugar. According to intake of food groups, significant differences were observed for five out of sixteen food groups. Spearman's correlation coefficients for energy, nutrients and food groups ranged from 0.43 (Ca) to 0.85 (soft drinks). Most Bland-Altman plots indicated broad limits of agreement. The differences between the two questionnaires can be explained by changes in the questionnaires, changes in the food composition databases used and random variation. Comparing differences between the questionnaires by maternal educational level, number of children and type of day care revealed minor differences. In conclusion, this study showed that at the group level there was reasonable comparability between the two questionnaires, except for carbohydrates, added sugar and some food groups. Moreover, there were moderate to high correlations for energy, nutrients and food groups.

Molecular Differentiation of Shigella Spp. from Enteroinvasive E. Coli.

A real-time polymerase chain reaction (PCR) assay, amplifying the genes encoding lactose permease (lacY) and invasion plasmid antigen H (ipaH), was run on 121 isolates phenotypically classified as Shigella spp., enteroinvasive Escherichia coli (EIEC), or EIEC O nontypable (ONT). The results were compared with data from a generic E. coli multiple-locus variable-number of tandem repeat analysis (MLVA) and a Shigella MLVA. The real-time PCR verified all Shigella spp. (n = 53) as Shigella (lacY negative) and all EIEC O121 (n = 15) and EIEC O124 (n = 2) as EIEC (lacY positive). However, the real-time PCR typed EIEC O164 as either EIEC (n = 2) or Shigella (n = 2) and, thus, was not suited for classifying this group of isolates. Interestingly, the majority (42/47, 89.4%) of the EIEC ONT were classified as Shigella (lacY negative) by the real-time PCR, and in nearly all cases, (92.9%, 39/42) data from both MLVA assays supported these findings. Overall, in 94.7% (114/121) of the isolates, the results from the real-time PCR were substantiated by the results from the MLVA assays. In conclusion, the real-time PCR assay was fast and accurate in differentiating Shigella spp. from EIEC, with the exception of the EIEC O164 group. This molecular assay was particularly pragmatic for the challenging EIEC ONT group.

Heat stress and protection from permanent acoustic injury in mice.

The inner ear can be permanently damaged by overexposure to high-level noise; however, damage can be decreased by previous exposure to moderate level, nontraumatic noise (). The mechanism of this "protective" effect is unclear, but a role for heat shock proteins has been suggested. The aim of the present study was to directly test protective effects of heat stress in the ear. For physiological experiments, CBA/CaJ mice were exposed to an intense octave band of noise (8-16 kHz) at 100 dB SPL for 2 hr, either with or without previous whole-body heat stress (rectal temperature to 41. 5 degrees C for 15 min). The interval between heat stress and sound exposure varied in different groups from 6 to 96 hr. One week later, inner ear function was assessed in each animal via comparison of compound action potential thresholds to mean values from unexposed controls. Permanent threshold shifts (PTSs) were approximately 40 dB in the group sound-exposed without previous heat stress. Heat-stressed animals were protected from acoustic injury: mean PTS in the group with 6 hr heat-stress-trauma interval was reduced to approximately 10 dB. This heat stress protection disappeared when the treatment-trauma interval surpassed 24 hr. A parallel set of quantitative PCR experiments measured heat-shock protein mRNA in the cochlea and showed 100- to 200-fold increase over control 30 min after heat treatment, with levels returning to baseline at 6 hr after treatment. Results are consistent with the idea that upregulation of heat shock proteins protects the ear from acoustic injury.

Quantitative studies of the non-productive binding of lysozyme to partially N-acetylated chitosans. Binding of large ligands to a one-dimensional binary lattice studied by a modified McGhee and von Hippel model.

This paper considers the non-productive (inhibitory) binding of chitosans to lysozyme from chicken egg white. Chitosans are linear, binary heteropolysaccharides consisting of 2-acetamido-2-deoxy-beta-D-glucose (GlcNAc; A-unit) and 2-amino-2-deoxy-beta-D-glucose (GlcN, D-unit). The active site cleft of lysozyme can bind six consecutive sugar residues in subsites named A-F, and specific binding of chitosan sequences to lysozyme occurs with A-units in subsite C. Chitosans with different fractions of A-units (FA) induced nearly identical changes in the 1H NMR spectrum of lysozyme upon binding, and the concentration of bound lysozyme could be determined. The data were analysed using a modified version of the McGhee and von Hippel model for binding of large ligands to one-dimensional homogeneous lattices. The average value of the dissociation constant for different sequences that may bind to lysozyme (K(ave)D) was estimated, as well as the number of chitosan units covered by lysozyme upon binding. K(ave)D decreased with increasing FA-values at pH 3 and 4.5, while the opposite was true at pH 5.5. Contributions from different hexamer sequences to K(ave)D of the chitosans were considered, and the data revealed interesting features with respect to binding of lysozyme to partially N-acetylated chitosans. The relevance of the present data with respect to understanding lysozyme degradation kinetics of chitosans is discussed.

Safety and tolerability of iodixanol in healthy volunteers with reference to two monomeric X-ray contrast media.

The low osmolar, non-ionic X-ray contrast media have shown a lower frequency of adverse events than the older ionic ones. In this study changes in routine clinical-chemical parameters in blood and urine, vital signs and adverse events were recorded in six groups of 10 healthy male volunteers receiving either iodixanol, a new non-ionic, dimeric X-ray contrast medium for general vascular use, or one of the two non-ionic, monomeric contrast media iopentol and iopamidol. Minor decreases were observed in the values for haemoglobin, haematocrit and erythrocytes 5 min and 3 days after injection of iodixanol. A minor increase was seen in platelets and total protein after 3 days. A transient increase in serum osmolality was seen 5 min after the injections of iopentol and iopamidol. This was not seen in any iodixanol group. The level of thyrotropin showed an increase in all groups at 3 days. It was back to normal within 21 days. No changes of clinical importance were seen regarding blood pressure, heart rate or ECG in any volunteer. No severe adverse events were reported. All events were of short duration, and of mild or moderate intensity. The results, however, may indicate a lower frequency of adverse events/discomfort after the administration of the dimeric iodixanol than the 2 monomeric contrast media iopentol and iopamidol.

Non-convulsive status epilepticus in the adult mentally retarded. Classification and role of benzodiazepines.

Non-convulsive status epilepticus (NCS) is rarely encountered and may appear with a psychiatric mask. As clouding of consciousness is the major ictal manifestation, the condition may easily be overlooked in the mentally retarded. We have studied 11 mentally retarded patients with NCS. Since NCS with a focal onset may have a generalized ictal EEG pattern, a classification of NCS solely based on the seizure classification may be misleading. In some patients, it is impossible, both clinically and on the basis of EEG recordings, to distinguish between continuous complex partial seizures and atypical absences. We therefore propose a revised classification of NCS based on the ictal EEG pattern and the epilepsy syndrome diagnosis (I) NCS in generalized epilepsy syndromes, (II) NCS in localization-related epilepsy, (a) with localized EEG features, (b) with generalized EEG features, and (c) with transitional EEG features, and (III) undetermined NCS. Four of our patients were classified as Group I, two as Group IIa, one as Group IIb, one as Group IIc, and three as Group III. Benzodiazepines at small or standard doses may be ineffective in terminating NCS, particularly in the Lennox-Gastaut Syndrome. The identification of trigger factors is essential. Drugs seemed to be the most important precipitants in our patients; in three, NCS was induced by recurrent rectal diazepam over-administration. This complication of rectal diazepam treatment in epilepsy has not been addressed previously.

Temporal bone histopathologic and genetic studies in Mohr-Tranebjaerg syndrome (DFN-1).

OBJECTIVE: To describe the temporal bone histopathologic and genetic abnormalities in a case of Mohr-Tranebjaerg syndrome. BACKGROUND: Mohr-Tranebjaezrg syndrome (DFN-1) is an X-linked, recessive, syndromic hearing loss, characterized by postlingual sensorineural hearing loss with onset in childhood, followed in adult life by progressive dystonia, spasticity, dysphagia, and optic atrophy. The syndrome is caused by mutations in the DDP (deafness/dystonia peptide) gene, which are thought to result in mitochondrial dysfunction with subsequent neurodegeneration. The temporal bone pathologic changes in this syndrome have not been reported. METHODS: Hearing loss developed in the patient at age 4, blindness at age 48, and dystonia at age 57. Genetic studies on peripheral blood showed a l51delT mutation in his DDP gene. He died at age 66. The right temporal bone was subjected to light microscopy and polymerase chain reaction-based analysis of the DDP gene sequence. RESULTS: There was near complete loss of spiral ganglion cells with loss of nearly all peripheral and central processes. Only 1,765 spiral ganglion cells remained (8.5% of mean normal for age). The organ of Corti (including hair cells), stria vascularis, and spiral ligament were preserved. There was also a severe loss of Scarpa's ganglion cells with preservation of vestibular hair cells. The population of geniculate and trigeminal ganglion cells appeared normal. Sequence analysis from temporal bone DNA showed the 15ldelT DDP gene mutation. CONCLUSION: Sensorineural hearing loss in Mohr-Tranebjaerg syndrome is the result of a postnatal, progressive, severe auditory neuropathy.

[Induced abortion. Reasons and contraceptive habits]. / Abortus provocatus. Begrundelser og kontraceptionsvaner.

An interview investigation comprizing 314 women referred for termination of pregnancy in Herlev Hospital revealed that the demographic conditions and the contraceptive habits for women applying for termination of pregnancy had not altered to any great extent since 1977-1978, when a similar investigation was carried out. Half of the women had not employed any form of contraception at the time of conception. The commonest reason stated was that the women had forgotten this. Thirty-six (11%) had not employed any form of contraception during the six months prior to conception. The group who had not employed contraception did not differ from the remainder as regards age and social status. 30% of the women had changed their method of contraception within the past six months. 17% had abandoned a relatively certain method in favour of the condom method because this method provides protection from sexually-transmitted diseases including AIDS. 32% of the single women had employed the condom method for this reason but, in half of these cases, the couple had forgotten to use the method during the coitus involved. As methods of reducing the number of terminations of pregnancy, information about the "morning-after" methods is primarily proposed. Offers of social assistance to applicants for training who constitute 27% of the women applying for termination of pregnancy, should be considered. The condom method should be supplemented by another method to increase the efficiency. On account of the serious somatic and physical problems involved in termination of pregnancy in very young women, efforts to prevent unwanted pregnancies should be continued in this particular age group.

[Is routine microscopy of tissue from induced abortions necessary?]. / Er rutinemaessig mikroskopi af vaev fra provokerede aborter nødvendig?

This study was performed in order to evaluate the importance of routine microscopy of tissue from legal abortions performed before the end of the 12th week of pregnancy. The tissues recovered from 580 abortions were examined carefully macroscopically before they were sent to microscopy. Five hundred and fifty-four samples (95.5%) were judged by macroscopic examination to contain the normal products of pregnancy. These all contained chorionic villi when examined histologically. Macroscopic examination was inconclusive in the remaining 26 samples (4.5%), 22 of these samples proved after all to contain chorionic villi when examined histologically. We conclude that histological examination of tissue from legal abortions is unnecessary in most cases if it is replaced by careful clinical examination of the sample, only to be followed by microscopy if the findings are uncertain.

A functional SUMO-interacting motif in the transactivation domain of c-Myb regulates its myeloid transforming ability.

c-Myb is an essential hematopoietic transcription factor that controls proliferation and differentiation of progenitors during blood cell development. Whereas sumoylation of the C-terminal regulatory domain (CRD) is known to have a major impact on the activity of c-Myb, no role for noncovalent binding of small ubiquitin-like modifier (SUMO) to c-Myb has been described. Based on the consensus SUMO-interacting motif (SIM), we identified and examined putative SIMs in human c-Myb. Interaction and reporter assays showed that the SIM in the in the transactivation domain of c-Myb (V(267)NIV) is functional. This motif is necessary for c-Myb to be able to interact noncovalently with SUMO, preferentially SUMO2/3. Destroying the SUMO-binding properties by mutation resulted in a large increase in the transactivation potential of c-Myb. Mutational analysis and overexpression of conjugation-defective SUMO argued against intramolecular repression caused by sumoylated CRD and in favor of SUMO-dependent repression in trans. Using both a myeloid cell line-based assay and a primary hematopoietic cell assay, we addressed the transforming abilities of SUMO binding and conjugation mutants. Interestingly, only loss of SUMO binding, and not SUMO conjugation, enhanced the myeloid transformational potential of c-Myb. c-Myb with the SIM mutated conferred a higher proliferative ability than the wild-type and caused an effective differentiation block. This establishes SUMO binding as a mechanism involved in modulating the transactivation activity of c-Myb, and responsible for keeping the transforming potential of the oncoprotein in check.

FLASH acts as a co-activator of the transcription factor c-Myb and localizes to active RNA polymerase II foci.

The c-Myb oncoprotein is a DNA-binding transcription factor with a key role in early stages of hematopoiesis. To expand our knowledge of partners cooperating with c-Myb, we performed a yeast two-hybrid screening with full-length c-Myb as bait. Here, we report FLICE-associated huge protein (FLASH)/CASP8AP2 as a novel Myb-interacting protein. We show that FLASH interacts with the DNA-binding domain of c-Myb and enhances c-Myb-dependent reporter activity and expression of endogenous c-Myb target genes. Chromatin immunoprecipitation assays revealed that FLASH and c-Myb both associate with the MYC promoter region as well as with the intronic enhancer of the c-Myb target gene ADA. Furthermore, siRNA knock-down of FLASH or c-Myb both result in a reduction of MYC and ADA expression. The co-activator effect is mediated through the C-terminal part of FLASH, which binds c-Myb. The FLASH-induced enhancement is comparable with the increase seen with the c-Myb co-activator p300. We find FLASH localized in discrete nuclear speckles in several cell lines, co-localized with c-Myb in active RNA polymerase II foci. These results imply a novel molecular mechanism of regulation of c-Myb activity. We propose that c-Myb cooperates with FLASH in foci associated with active RNA polymerase II, leading to enhancement of Myb-dependent gene activation.

[Norwegian fibromyalgia epidemic--its rise or possible decline. What is the trend based on disability statistics?]. / Den norske fibromyalgi-epidemiens vekst--og mulige fall. Hva viser uførestatistikken?

The Norwegian National Insurance Scheme (Folketrygden) offers universal coverage to all inhabitants between 16 and 67 years with respect to disability pension. During the 1980s, the number of new disability-pensioners increased rapidly. In 1991, 8.5% of the population at risk received this pension. So called "diffuse" conditions in the musculoskeletal system accounted for a large proportion of new cases. Myalgia/fibromyalgia became a major reason for disability pension. In 1989 more than 7% of the new cases had this diagnosis. The parliament (Stortinget) passed controversial amendments to the National Insurance Acts in 1991 and 1995 which restricted the criteria for obtaining a disability pension. At present the law demands that "a scientific concept of disease" should be applied in these matters. The numbers of new disability-pensioners decreased significantly during the period 1989-1993. The figures from 1994 and 1995 perhaps show a new upward trend, including also "diffuse" diagnoses like fibromyalgia. The use of "diffuse conditions" as a cause for disability pension is discussed in light of the official request for a scientifically justified diagnosis.

Micropuncture measurements of interstitial fluid pressure in the rat periodontal ligament.

The interstitial fluid pressure (IFP) or so-called tissue pressure in the periodontal ligament (PDL) has been suggested to play a role in tooth position and affect periodontal blood flow. However, few investigators have actually measured the tissue pressure in the PDL and measurements in the PDL of rat continuously erupting incisors are lacking. The present study was aimed to develop the micropuncture technique for measurements of IFP in the PDL. The PDL of rat incisors was exposed by partial removal of the crown, the coronal pulp was removed. Viewed through a stereomicroscope, a cavity was drilled until the vessels of the PDL were visible through a thin layer (20 microns) of root cement. The cavity was covered with saline and the PDL punctured with glass micropipettes, diameter 2-6 microns, through tiny cracks of the root cement. The periodontal tissue pressure (IFP) was measured by connecting the micropipette to a servocontrolled counter-pressure pump system (Wiederhielm 1964). Mean IFP in 22 Sprague-Dawley rats was 15.2(SD +/- 3.6) mmHg and simultaneously measured systemic arterial pressure (PA) averaged 125 mmHg. Intravenous administration of an antiflogisticum, indomethacin (15 rats), did not change IFP significantly (mean 14.3, SD +/- 4.2 mmHg). Increased blood volume in the PDL, induced by bilateral clamping of the jugular veins, resulted in an immediate rise in IFP, whereas reduced blood volume due to cardiac arrest arrest resulted in an abrupt fall in IFP, indicating low compliance in the rat PDL. It is concluded that the measurements with the present micropuncture technique are relatively atraumatic and thus it seems to be a reliable method for recordings of IFP in the PDL.

Effect of experimental traumatic occlusion on blood flow in the temporomandibular joint of the rat.

Fluorescent microspheres (FM) were used to visualize and semi-quantify blood flow in the temporomandibular joint (TMJ) during experimental unilateral traumatic occlusion of the maxillary and mandibular molar teeth in 30 young rats. At different postoperative observation periods varying from 1 to 30 days FM were injected systemically, and the number of FM were counted in serial sections from the TMJ in a fluorescent microscope. Blood flow was related to the number of FM found in the fibrous connective tissue and bony condyle of the TMJ. A percentage increase in blood flow was found at 15 to 20 days on the experimental side, compared with the contralateral side. Furthermore, there was an increase in blood flow in both TMJs in the experimental animals compared with an unoperated control material of 10 animals. The study thus indicates that a local unilateral occlusal trauma initiates blood flow responses not only unilaterally but also in the TMJ on both sides in rats.

Increased pressure in venous sinusoids during decongestion of rat nasal mucosa induced by adrenergic agonists.

The haemodynamic effects of sympathetic agonists causing decongestion of the nasal mucosa have been investigated in rats. Access to mucosa was obtained from the dorsal side through a small cavity drilled in the nasal bone. The pressures in the venous sinusoids and in the interstitial fluid of nasal mucosa were recorded by micropuncture technique. The local red cell flux (LDF) was monitored by laser Doppler flowmetry, and the blood volume in the mucosa was measured by radio-labelled erythrocytes and albumin. In control rats the tissue blood volume was 0.25 +/- 0.03 g (g wet wt)-1. The interstitial fluid pressure (IFP) was 2.4 +/- 0.6 mmHg and the average blood pressure in venous sinusoids (Ps) was 12.8 +/- 2.7 mmHg. After topical application of noradrenaline (NA) the local blood volume was reduced to 0.12 +/- 0.03 g g-1. Ps was increased to 18.0 +/- 4.0 mmHg, whereas IFP was maintained and LDF was reduced to 40.4% of control, indicating a greater rise in post than in presinusoid vascular resistance. Blocking of both alpha 1 and alpha 2-receptors by phentolamine caused a rise in mucosa blood volume and in LDF by 16 and 20% of control, respectively. Ps increased significantly to 15.2 +/- 3.3 mmHg. Specific stimulation or blocking of alpha 1-receptors by phenylephrine or prazosin induced similar or slightly smaller vascular responses than NA or phentolamine. The effects of the specific alpha 2-agonist (clonidine) or antagonist (yohimbine) on rat mucosa were small, indicating a domination of the alpha 1-receptors. Thus, application of NA caused a rise in blood pressure in the venous sinusoids of nasal mucosa. As LDF fell simultaneously, the reduced blood volume must be due to an increased tone in the muscular wall of venous sinusoids.