Analysis of the small chromosomal Prionium serratum (Cyperid) demonstrates the importance of reliable methods to differentiate between mono- and holocentricity.

For a long time, the Cyperid clade (Thurniceae-Juncaceae-Cyperaceae) was considered a group of species possessing holocentromeres exclusively. The basal phylogenetic position of Prionium serratum (Thunb.) Drège (Thurniceae) within Cyperids makes this species an important specimen to understand the centromere evolution within this clade. In contrast to the expectation, the chromosomal distribution of the centromere-specific histone H3 (CENH3), alpha-tubulin and different centromere-associated post-translational histone modifications (H3S10ph, H3S28ph and H2AT120ph) demonstrate a monocentromeric organisation of P. serratum chromosomes. Analysis of the high-copy repeat composition resulted in the identification of two centromere-localised satellite repeats. Hence, monocentricity was the ancestral condition for the Juncaceae-Cyperaceae-Thurniaceae Cyperid clade, and holocentricity in this clade has independently arisen at least twice after differentiation of the three families, once in Juncaceae and the other one in Cyperaceae. In this context, methods suitable for the identification of holocentromeres are discussed.

Activity and inactivity of moth sex chromosomes in somatic and meiotic cells.

Moths and butterflies (Lepidoptera) are the most species-rich group of animals with female heterogamety, females mostly having a WZ, males a ZZ sex chromosome constitution. We studied chromatin conformation, activity, and inactivity of the sex chromosomes in the flour moth Ephestia kuehniella and the silkworm Bombyx mori, using immunostaining with anti-H3K9me2/3, anti-RNA polymerase II, and fluoro-uridine (FU) labelling of nascent transcripts, with conventional widefield fluorescence microscopy and 'spatial structured illumination microscopy' (3D-SIM). The Z chromosome is euchromatic in somatic cells and throughout meiosis. It is transcriptionally active in somatic cells and in the postpachaytene stage of meiosis. The W chromosome in contrast is heterochromatic in somatic cells as well as in meiotic cells at pachytene, but euchromatic and transcriptionally active like all other chromosomes at postpachytene. As the W chromosomes are apparently devoid of protein-coding genes, their transcripts must be non-coding. We found no indication of 'meiotic sex chromosome inactivation' (MSCI) in the two species.

A novel frameshift variant in the CADASIL gene NOTCH3: pathogenic or not?

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) represents the most common monogenic cause of adult-onset ischemic stroke and vascular dementia. It is caused by heterozygous missense mutations in the NOTCH3 gene, encoding a transmembrane receptor protein on vascular smooth muscle cells. Classical CADASIL mutations affect conserved cysteine residues of the Notch3 protein. By contrast, the role of non-canonical genetic variation in NOTCH3, in particular of variants causing a hypomorphic Notch3 protein, is subject to an ongoing scientific debate. In this context, we here report a novel NOTCH3 frameshift variant in exon 18 (NM_000435.2: c.2853_2857delTCCCG), causing a frameshift and introducing a premature stop codon, which was detected in a 43-year-old woman and her father. Both carriers of the variant were carefully evaluated, including serial follow-up in the index. Neither clinical nor imaging features provided convincing evidence for a classical CADASIL phenotype, thus reinforcing the concept of hypomorphic NOTCH3 variants most likely not being causative for CADASIL. Our finding, which is discussed in the light of the published literature, has practical implications for interpreting results of NOTCH3 molecular genetic testing as well as patient counseling.

Anti-phosphorylated histone H2AThr120: a universal microscopic marker for centromeric chromatin of mono- and holocentric plant species.

Based on the analysis of 20 different monocot and eudicot species, we propose that the centromeric distribution of the phosphorylated histone H2AThr120 is evolutionary highly conserved across species with mono- and holocentric chromosomes. Therefore, antibodies recognizing the phosphorylated threonine 120 of the histone H2A can serve as a universal marker for the cytological detection of centromeres of mono- and holokinetic plant species. In addition, super resolution microscopy of signals specific to the centromere-specific histone H3 variant CENH3 and to H2AThr120ph revealed that these histone variants are incorporated into different nucleosomes, which form distinct, partly intermingled chromatin domains. This specific arrangement of both histone variants suggests different centromeric functions during the cell cycle.

No neocentric activity on Aegilops markgrafii chromosome E.

Classical neocentromeres, proven mainly in Poaceae species, cause meiotic drive in higher plants. Here I present data indicating that the morphological stretching of a chromatin domain at the long arm of Aegilops markgrafii chromosome E during meiosis is not due to a supposed neocentric activity. The stretching is visible at the alien univalent in metaphase I and anaphase I pollen mother cells of Triticum aestivum--Ae. markgrafii monosomic addition line E and at one univalent of the 49-chromosome-containing plants containing the haploid Ae. markgrafii genome added to wheat. The absence of both centromeric histone variant CENH3 and attachment of spindle fibers, proven by immunostaining with antibodies against CENH3 and tubulin, respectively, revealed that the univalent elongation of chromosome E is not caused by a classical plant neocentromere.

SMC proteins and their multiple functions in higher plants.

In plants as in other eukaryotes, SMC (Structural Maintenance of Chromosome) protein complexes and proteins interacting with them are essential for sister chromatid cohesion, chromosome condensation, DNA repair and recombination. The presence of paralogous genes for various components of the different SMC complexes allowed diversification of their biological functions during evolution of higher plants. Here I summarize the multiple functions of plant SMC complexes and some of the particularities these proteins show in comparison to those of other organisms.

Nuclear DNA content and nuclear and cell volume are positively correlated in angiosperms.

Volumes of flow sorted nuclei were analyzed from two highly endopolyploid (diploids with endopolyploid tissues) species (Arabidopsis thaliana and Barbarea stricta), from a less endopolyploid species (Allium cepa) and from two non-endopolyploid species (Chrysanthemum multicolor and Fritillaria uva-vulpis). Intraspecific as well as interspecific comparisons revealed a highly positive correlation (r > 0.99) between DNA content and nuclear volume. No significant differences between expected and measured nuclear volumes were noted indicating that chromatin packing is not increased with increasing DNA content in the tested plant species. In epidermis cells of A. thaliana, A. cepa and Ch. multicolor, a lower (r between 0.6 and 0.7) but significant positive correlation between nuclear volume and cell volume was found. This correlation is compatible with the hypothesis that endopolyploidization (EP = consecutive replication cycles not separated by nuclear and cell divisions) might speed up the growth of endopolyploid species and compensate for small genome size.

DNA damage processing and aberration formation in plants.

Various types of DNA damage, induced by endo- and exogenous genotoxic impacts, may become processed into structural chromosome changes such as sister chromatid exchanges (SCEs) and chromosomal aberrations. Chromosomal aberrations occur preferentially within heterochromatic regions composed mainly of repetitive sequences. Most of the preclastogenic damage is correctly repaired by different repair mechanisms. For instance, after N-methyl-N-nitrosourea treatment one SCE is formed per >40,000 and one chromatid-type aberration per approximately 25 million primarily induced O6-methylguanine residues in Vicia faba. Double-strand breaks (DSBs) apparently represent the critical lesions for the generation of chromosome structural changes by erroneous reciprocal recombination repair. Usually two DSBs have to interact in cis or trans to form a chromosomal aberration. Indirect evidence is at hand for plants indicating that chromatid-type aberrations mediated by S phase-dependent mutagens are generated by post-replication (mis)repair of DSBs resulting from (rare) interference of repair and replication processes at the sites of lesions, mainly within repetitive sequences of heterochromatic regions. The proportion of DSBs yielding structural changes via misrepair has still to be established when DSBs, induced at predetermined positions, can be quantified and related to the number of SCEs and chromosomal aberrations that appear at these loci after DSB induction. Recording the degree of association of homologous chromosome territories (by chromosome painting) and of punctual homologous pairing frequency along these territories during and after mutagen treatment of wild-type versus hyperrecombination mutants of Arabidopsis thaliana, it will be elucidated as to what extent the interphase arrangement of chromosome territories becomes modified by critical lesions and contributes to homologous reciprocal recombination. This paper reviews the state of the art with respect to DNA damage processing in the course of aberration formation and the interphase arrangement of homologous chromosome territories as a structural prerequisite for homologous rearrangements in plants.

The effects of light therapy on mini-mental state examination scores in demented patients.

BACKGROUND: Preliminary evidence suggests that demented patients may experience beneficial effects of light therapy. The authors tested whether bright light therapy (BLT) is capable of improving cognitive functions in patients with Alzheimer-type dementia (AD) or vascular dementia (VD). METHODS: Twenty-three patients with AD or VD were randomly assigned to either evening BLT or dim light therapy (DLT). Effects of light therapy on cognitive functions were assessed before and after light therapy using Mini-Mental State Examination (MMSE) scores. Body temperature rhythm (BTR) was additionally recorded pre- and posttreatment. RESULTS: Irrespective of their diagnosis, patients treated with BLT (p =.0012) but not with DLT (p =.73) showed a statistically significant increase in MMSE total scores after light therapy. Evening BLT simultaneously induced a significant phase delay of 56 min on BTR (p =.025). CONCLUSION: Our preliminary results suggest that short-term evening BLT may exert beneficial effects on cognitive functioning in patients with dementia.

Effects of phototherapy on pressure ulcer healing in elderly patients after a falling trauma. A prospective, randomized, controlled study.

BACKGROUND: The effects of infrared and red pulsed monochromatic light, with varied pulsations and wavelengths, on the healing of pressure ulcers were evaluated in this prospective, randomized, controlled study. METHODS: Elderly patients (> or =65 years) with Stage 2 or 3 skin ulcers were enrolled and assigned to one of two groups. Both groups were given the same standard ulcer therapy. One group was also given phototherapy with pulsed monochromatic infrared (956 nm) and red (637 nm) light. Treatments lasted 9 min each time using a regimen with pulse repetition frequency varied between 15.6 Hz and 8.58 kHz. Patients were followed for 10 weeks or until the ulcer was healed, whichever occurred first. The ulcer surface area was traced weekly. RESULTS: Patients treated with pulsed monochromatic light had a 49% higher ulcer healing rate, and a shorter time to 50% and to 90% ulcer closure compared with controls. Their mean ulcer area was reduced to 10% after 5 weeks compared with 9 weeks for the controls. CONCLUSION: The results are encouraging as pulsed monochromatic light increased healing rate and shortened healing time. This will positively affect the quality of life in elderly patients with pressure ulcer.

The influence of local heating on skin microcirculation in pressure ulcers, monitored by a combined laser Doppler and transcutaneous oxygen tension probe.

In a prospective, controlled study the skin microcirculation and transcutaneous oxygen tension were evaluated in 28 elderly patients (85 +/- 0.8 years) with grade 2 and 3 pressure ulcer. Laser Doppler fluxmetry and transcutaneous oxymetry techniques were used together in a combined probe to simultaneously evaluate the influence of heat stress at 44 degrees C, at the same time and in the same area of the ulcer edge. Total skin microcirculation was already increased at rest before heating, and increased even more during a fast and then a more gradually heat-induced hyperaemia response compared with undamaged skin. The increase showed a biphasic manner. On the other hand, the oxygen diffusibility from the capillaries to the skin surface was significantly reduced, compared with undamaged skin. One hypothesis may be that in ageing skin the main part of the increased skin microcirculation in a pressure ulcer were passing through thermoregulatory vessels in subpapillary tissue layers. Interactions between the increased blood flow and metabolic activity at the ulcer edge might be oxygen-consuming, leading to reduced oxygen content passing through the capillaries and contributing to tissue ischaemia. Disturbances of the local skin microcirculation and tissue oxygenation will influence the ulcer healing rate and may affect the healing progress.

[Letting family participate in care. A study on relatives' experience with a palliative geriatric care department]. / Att dela med sig av vården till anhöriga. En studie om anhörigas upplevelse på en palliativ geriatrisk vårdavdelning.

An evaluation was made of relatives' experiences of the care of the patient and the treatment of themselves at a geriatric ward where elderly patients dying of cancer were nursed. A questionnaire was sent to 100 relatives after they had agreed on telephone to participate. 86 relatives answered the questions. The results shows that the majority of the relatives were satisfied with the care of the patient and the treatment of themselves as relatives. Most of them thought that they had been able to influence the care of their next-to-kin. However, elderly ladies and younger men were uncertain whether they had had the possibility to influence the care. Most of the relatives had been given the opportunity to talk enough to the doctor, the nurses and the social-worker. However; they required a more active approach from the staff in the ward. They also desired more information about the patient's disease, prognosis and medical treatment. The findings of this study suggest that relatives need information repeated at several occasions in order to understand the message. They also need support and invitation to get the courage to ask questions and to participate in the care of their dying next-of-kin. We need deepen our knowledge of what kind of information the relatives demands and what information needs to be repeated. Evaluations of routines to make contact easier between patients, their relatives and hospital staff should be done continuously.

Analysis of the measurement of four wound variables in elderly patients with pressure ulcers.

The suitability of four different methods for measuring pressure ulcer size was investigated during 10 consecutive weeks in a group of 11 elderly patients. The methods used were: 1. Volume measurement by filling the wound cavity with a gel 2. area measurement, by tracing the wound boundary outlined on transparent film with a digital planimeter 3. perimeter measurement, by tracing the wound boundary outlined on transparent film with a digital measuring pen 4. depth measurement using a millimeter probe. The mean values of the rate parameters for volume, area, perimeter, and depth were approximately in the proportions 3:2:1:1, which mirrors the inherent geometric dimensions of the corresponding variables. The methods used for measuring perimeter and depth were less reliable than the methods used for measuring volume and area. Thus, the use of both volume and area measurements in recommended. The former is best suited for deep ulcers and the latter for broad, irregularly shaped ulcers.

Analysis of arterial flowmotion in spinal cord injured and elderly subjects in an area at risk for the development of pressure sores.

Flowmotion (blood flow changes due to vasomotion) in the skin over a risk area for pressure sores, the sacrum, and a non-risk area, the gluteus muscle, was evaluated by laser Doppler fluxmetry during resting conditions and post-occlusive reactive hyperaemia (PRH) response. Measurements were made in healthy younger subjects and in two risk groups for pressure sores-spinal cord injured (SCI) and the elderly. The SCI were divided into two subgroups, one with and one without distinct flowmotion seen on the original recordings over the sacrum. The Prony spectral line estimation (PSLE) method was used to determine the power spectrum of the flowmotion activity. During the PRH, flowmotion frequencies were found in two separated bands, 5.4-6.6 cpm (cycles min-1) and 7.8-9.0 cpm. In the subgroup without distinct flowmotion, the PSLE method found flowmotion frequencies similar to the other groups. During the PRH, the flowmotion power pattern over the sacrum was similar within all groups, but the power was extremely low in one subgroup of SCI subjects. To conclude, flowmotion (vasomotion) was present in the two skin areas and increased during the PRH response, reaching a maximum within 50 s. This study shows that the flowmotion frequencies might be locally driven, whereas the power might be centrally mediated. Disturbances in microcirculatory flowmotion can be a part of the mechanisms leading to skin ischaemia and pressure sores. A new method has been developed for analysing differences in flowmotion behaviour such that statistical comparisons can be made.

The effects of pressure and shear on skin microcirculation in elderly stroke patients lying in supine or semi-recumbent positions.

The effects of external pressure and shear on the skin microcirculation over the sacral area, which is known as a high risk area for pressure sore formation, were studied in 30 elderly patients. The skin blood cell flux (SBF) was measured using the laser Doppler technique, with the patient first at rest in lateral position, then lying for 30 minutes in supine or semi-recumbent 45 degrees position, and finally in lateral position. Elderly high-risk patients (G2), most of them more than two years post-stroke, had a lower body mass index and a reduced sacral skin-fold compared with non-risk patients (G1). The SBF in G2 decreased 28% in supine and 14% in 45 degrees position, whereas the SBF in G1 increased 35% in supine and 13% in 45 degrees position. Spontaneous movements up to seven times per 30 minutes were registered, even during sleep, and were evident by direct observation of the recorded charts as a temporary SBF increase. The risk for skin ischaemic damage over the sacral area of elderly risk patients was evident in both positions, especially with the patients lying in supine position. When increasing the upper body slope in G2 from horizontal to 45 degrees, an inability to recover a satisfactory blood supply after the ischaemic insult was found. Discomfort from compressive and shear forces initiates changes in posture, even in elderly patients prone to tissue breakdown. Occasional relief of pressure was in most patients followed by temporary increase in skin blood flow with concomitant temperature increase. This most probably protected them from developing skin lesions.

Skin microcirculatory and thermal changes in elderly subjects with early stage of pressure sores.

Elderly subjects are prone to develop pressure sores over the sacrum area mainly due to external pressure and shear effects which negatively affect the skin microcirculation. The aim of the study described here was to measure skin microcirculatory and thermal changes in twelve elderly patients with an early stage of pressure sore and in ten elderly subjects without pressure sore. The total skin microcirculation at a damaged risk area, the sacrum, and a reference area of undamaged skin over the gluteal region, was evaluated using the laser Doppler fluxmetry. The nutritive transport of small solutes, characterized by the transcapillary exchange of sodium fluorescein, was evaluated using the fluorescein flowmetry technique. The skin temperature was measured with a thermoelement. The skin microcirculation at the area with an early stage of pressure sore was significantly higher than at undamaged skin as measured by both techniques. The total microcirculatory blood flow increased > 16 times (P < 0.001) and the nutritive transport approximately 5 times (P < 0.01) compared to the reference value, and the calculated blood flow of subpapillary tissue layers increased 17-19 times (P < 0.001). However, the skin temperatures in damaged and undamaged skin did not differ significantly. These results show an increased skin microcirculation in the early stage of pressure sores but no increase in skin temperature. The more strongly increased skin blood flow in subpapillary tissue layers effectively conducts away the heat caused by the damage and the increased metabolic activity.

Repeated DNA sequences of Aegilops markgrafii (Greuter) Hammer var. markgrafii: cloning, sequencing and analysis of distribution in Poaceae species.

Total DNA of Aegilops markgrafii (Greuter) Hammer var. markgrafii was shot gun cloned. From all the recombinants containing repetitive sequences 1-2% hybridized preferentially with the Ae. markgrafii genome and were almost absent in wheat. The cloned sequences are disperse distributed over the Aegilops chromosomes and show the typical features of eukaryotic repetitive DNA. Five specific probes were tested for their applicability in a screening program on 68 Poaceae accessions.

C-banding pattern and polymorphism of Aegilops caudata and chromosomal constitutions of the amphiploid T. aestivum - Ae. caudata and six derived chromosome addition lines.

C-banding patterns were analysed in 19 different accessions of Aegilops caudata (= Ae. markgrafii, = Triticum dichasians) (2n = 14, genomically CC) from Turkey, Greece and the USSR, and a generalized C-banded karyotype was established. Chromosome specific C-bands are present in all C-genome chromosomes, allowing the identification of each of the seven chromosome pairs. While only minor variations in the C-banding pattern was observed within the accessions, a large amount of polymorphic variation was found between different accessions. C-banding analysis was carried out to identify Ae. caudata chromosomes in the amphiploid Triticum aestivum cv 'Alcedo' - Ae. caudata and in six derived chromosome addition lines. The results show that the amphiploid carries the complete Ae. Caudate chromosome complement and that the addition lines I, II, III, IV, V and VIII carry the Ae. caudata chromosome pairs B, C, D, F, E and G, respectively. One of the two SAT chromosome pairs (A) is missing from the set. C-banding patterns of the added Ae. caudata chromosomes are identical to those present in the ancestor species, indicating that these chromosomes are not structurally rearranged. The results are discussed with respect to the homoeologous relationships of the Ae. caudata chromosomes.

Hypotension as a risk factor for the development of pressure sores in elderly subjects.

The systolic (sBP), diastolic (dBP) and mean (mBP) blood pressure were measured in hospitalized geriatric patients with (n = 30) and without (n = 100) pressure sores. A more intensive investigation was carried out in 18 elderly hospitalized geriatric patients without pressure sores, who were compared with 10 healthy young and 10 healthy elderly subjects. In these groups, the blood pressure was compared to the skin blood cell flux (SBF) as evaluated by the laser Doppler technique, at rest and during the post-occlusive reactive hyperaemia (PRH) response in the sacral area which is a high risk area for pressure sores. The patients with pressure sores had a significantly lower sBP (130 +/- 17 mmHg) compared to patients without pressure sores (140 +/- 20 mmHg). A significant correlation between a low resting SBF and a low mBP was found in all groups. An impaired PRH response corresponded to a low sBP in the two elderly groups. No correlation to dBP was seen at all. It is concluded that low mBP and sBP in elderly hospitalized patients were significantly correlated to a low resting SBF and a impaired reactivity of the skin microcirculation, respectively, and a low sBP was associated with pressure sores. Measurements of the systemic BP are of importance when classifying patients at risk of pressure sores.

Evaluation of the dynamic cutaneous post-ischaemic hyperaemia and thermal response in elderly subjects and in an area at risk for pressure sores.

The response of skin blood cell flux (SBF) to locally applied pressure was evaluated with the laser-Doppler technique in the areas of the sacrum and the gluteus maximus muscle of geriatric patients and healthy young and elderly subjects. The SBF over the sacrum stopped at a lower external skin pressure than over the gluteus muscle in all groups studied (P less than 0.05-0.001). The SBF at rest was lower among geriatric patients and over the sacrum, with men showing the lowest value (P less than 0.001). The initial slope of the SBF curve and the peak SBF during the post-occlusive reactive hyperaemia (PRH) were lower among geriatric patients compared to younger healthy subjects (P less than 0.05-0.01) over both areas studied. During the last part of the PRH response rhythmic oscillations started, known as vasomotion. The mean skin temperature at rest was higher over the sacrum (P less than 0.001) than over the gluteus area, but no difference was found between the groups. The temperature increase during the PRH was larger over the gluteus muscle (P less than 0.01) than over the sacrum area in the healthy subjects. Among the patients the temperature increase was larger over the gluteus only during the first half of the PRH, after which the temperature increased most over the sacrum. It is concluded that SBF and skin temperature show different responses in microvascular reactivity to external pressure in elderly vs. younger subjects and also between sexes. The described technique may be useful in clinical practice for predicting the risk of pressure sores in different areas and subjects.