Skin donations after suicide by acute poisoning.

The demand for skin tissue allografts to treat burns and other types of injuries increases each year to the extent that categories of donors formerly deemed "unsuitable", such as victims of suicide by polytrauma or poisoning, are now considered. Patients who died by ingestion of/exposure to toxic substances can be accepted as tissue donors after assessment of graft safety to rule out any risks of transferring toxic substances to the recipient. A cadaveric skin donation was obtained from a 57-year-old woman who died from intoxication after ingesting colchicine tablets (0.2 mg/kg). To determine the safety of cadaveric skin allografts, high-performance liquid chromatography-mass spectrometry (LC-MS/MS) was used to identify and quantify colchicine in procured skin. Results revealed that colchicine concentrations were lower than the instrument limit of detection (LOD) of 0.5 ng/mg both in epidermis and dermis. Cell viability assessed through the MTT ([3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]) test was within standard limits. Thanks to accurate tests performed, which are routinely applied also in clinical diagnostics and forensic toxicology, it was possible to ascertain the safety and suitability of skin tissue for donation.

Prevalence of sealant in an Italian paediatric population: an epidemiological study.

AIM: The use of sealants is an effective measure for the prevention of pit and fissure caries in children and it has been well documented by several studies In order to plan and establish a preventive national programme, it is important to know the epidemiological pattern in an Italian paediatric population, correlated to the risk of caries, DMFT and other sociodemographic factors. METHODS: This study was conducted on 2,442 children aged between 6 and 12 years attending the paediatric dentistry department of the University of L'Aquila, Italy. In addition to the oral examination, a questionnaire was administered on bad habits and the family perception of sealing. For descriptive analysis, the sample was stratified into two groups based on the presence/absence of at least one tooth with sealant. The differences between discrete and nominal variables, reported as absolute and percentage frequencies, were assessed by applying the χ2 test or the Fisher's exact test, as appropriate. Continuous variables were expressed in terms of mean values and their standard deviation(±DS) and the differences between the two groups under consideration were analysed through Student's t-test. The tests used are two-way and a significance level of 5% was applied. The statistical analysis was carried out using the statistical package STATA/IC 15.0 (StataCorp LLC, Texas, USA). CONCLUSION: The application of sealants to healthy occlusal surfaces is the best aid in preventing the development of caries in these areas, and this is especially important in childhood and adolescence, when the incidence of this pathology is particularly high.

Prevalence of molar incisor hypomineralization among school children in Rome, Italy.

Molar incisor hypomineralization (MIH) is a highly prevalent condition associated with increased caries experience, dental pain and treatment need. Aim of this study was to determine the prevalence and severity of MIH in a group of 7-8 years old primary school children living in Rome, Italy; and to assess the association with caries experience and possible perinatal risk factors. A survey has been conducted in the city of Rome, between April 2019 and March 2020 with a total of 49 primary schools and 176 2nd grade primary school classes and a total of 3611 children being involved. Of these, a subset of 346 children of 21 primary schools was selected for the epidemiological investigation. The prevalence of MIH was of 18.2%, with girls showing twice the probability of being subject to a mild-severe condition. Molar location was present in 71.4%, while location on both molar plus incisor was present in 28.6% of cases. The mean DMFT was 0.44 ± 0.78, "D" was 0.17 ± 0.58; the mean dmft was 1.7 ± 2.56, "d" was 1.32 ± 2.21. Female gender, caries experience, insufficient oral hygiene were risk factors. The incidence of MIH is increasing in the pediatric population. Knowledge about diagnosis and treatment options should be disseminated among dental professionals.

How to define a quadruple aim framework to assess value in critical pathway of the patients with Clostridioides difficile infection.

OBJECTIVE: The study aims to define the set of Key Performance Indicators (KPIs) required to assess the Value delivered by managing patients with Clostridioides difficile infection through a Critical Pathway. We used the quadruple aim Value-Based approach, and we validated the set of KPIs with the Delphi method. MATERIALS AND METHODS: The study focuses on patients on board a Critical Pathway on Clostridioides difficile Infection and targeted towards a Fecal Microbiota Transplantation (FMT). FMT has been used to successfully treat recurrent Clostridium difficile infection. A two-round e-Delphi survey collecting data was conducted in 2019-2020 to validate the Value-Based evaluation tool. The Value-Based criteria taken into account are Clinical Outcomes, Experience of Care, Per-capita cost, Physician's burnout. RESULTS: The two rounds led to the validation of 50 items, and four primary clinical outcomes (Mortality rate, length of stay, readmission and complications related to the illness). CONCLUSIONS: The evaluation tool included is validated in its totality and can provide a comprehensive overview of the Value created by the Critical pathway for patients with Clostridioides difficile. We can extend the approach illustrated in this study can also to evaluate other Critical pathways.

The anatomy of A-, B-, and Z-DNA.

Recent advances in DNA synthesis methods have made it possible to carry out single-crystal x-ray analyses of double-stranded DNA molecules of predetermined sequence, with 4 to 12 base pairs. At least one example has been examined from each of the three known families of DNA helix: A, B, and Z. Each family has its own intrinsic restrictions on chain folding and structure. The observed solvent positions in these crystal structures have confirmed earlier fiber and solution measurements, and have led to proposals explaining the transitions from B to A and from B to Z helices. Prospects are improving for an understanding of the mode of bending of DNA in chromatin, and the way in which specific DNA sequences are recognized by drug molecules and repressor proteins.

Cortical recruitment and functional dynamics in postural control adaptation and habituation during vibratory proprioceptive stimulation.

OBJECTIVE: Maintaining upright posture is a complex task governed by the integration of afferent sensorimotor and visual information with compensatory neuromuscular reactions. The objective of the present work was to characterize the visual dependency and functional dynamics of cortical activation during postural control. APPROACH: Proprioceptic vibratory stimulation of calf muscles at 85 Hz was performed to evoke postural perturbation in open-eye (OE) and closed-eye (CE) experimental trials, with pseudorandom binary stimulation phases divided into four segments of 16 stimuli. 64-channel EEG was recorded at 512 Hz, with perturbation epochs defined using bipolar electrodes placed proximal to each vibrator. Power spectra variation and linearity analysis was performed via fast Fourier transformation into six frequency bands (Δ, 0.5-3.5 Hz; θ, 3.5-7.5 Hz; α, 7.5-12.5 Hz; ß, 12.5-30 Hz; [Formula: see text], 30-50 Hz; and [Formula: see text], 50-80 Hz). Finally, functional connectivity assessment was explored via network segregation and integration analyses. MAIN RESULTS: Spectra variation showed waveform and vision-dependent activation within cortical regions specific to both postural adaptation and habituation. Generalized spectral variation yielded significant shifts from low to high frequencies in CE adaptation trials, with overall activity suppressed in habituation; OE trials showed the opposite phenomenon, with both adaptation and habituation yielding increases in spectral power. Finally, our analysis of functional dynamics reveals novel cortical networks implicated in postural control using EEG source-space brain networks. In particular, our reported significant increase in local θ connectivity may signify the planning of corrective steps and/or the analysis of falling consequences, while α band network integration results reflect an inhibition of error detection within the cingulate cortex, likely due to habituation. SIGNIFICANCE: Our findings principally suggest that specific cortical waveforms are dependent upon the availability of visual feedback, and we furthermore present the first evidence that local and global brain networks undergo characteristic modification during postural control.

Ordered water structure around a B-DNA dodecamer. A quantitative study.

The crystal structure of the double-helical B-DNA dodecamer of sequence C-G-C-G-A-A-T-T-C-G-C-G has been solved and refined independently in three forms: (1) the parent sequence at room temperature; (2) the same sequence at 16 K; and (3) the 9-bromo variant C-G-C-G-A-A-T-TBrC-G-C-G at 7 degrees C in 60% (v/v) 2-methyl-2,4-pentanediol. The latter two structures show extensive hydration along the phosphate backbone, a feature that was invisible in the native structure because of high temperature factors (indicating thermal or static disorder) of the backbone atoms. Sixty-five solvent peaks are associated with the phosphate backbone, or an average of three per phosphate group. Nineteen other molecules form a first shell of hydration to base edge N and O atoms within the major groove, and 36 more are found in upper hydration layers. The latter tend to occur in strings or clusters spanning the major groove from one phosphate group to another. A single spermine molecule also spans the major groove. In the minor groove, the zig-zag spine of hydration that we believe to be principally responsible for stabilizing the B form of DNA is found in all three structures. Upper level hydration in the minor groove is relatively sparse, and consists mainly of strings of water molecules extending across the groove, with few contacts to the spine below. Sugar O-1' atoms are closely associated with water molecules, but these are chiefly molecules in the spine, so the association may reflect the geometry of the minor groove rather than any intrinsic attraction of O-1' atoms for hydration. The phosphate O-3' and O-5' atoms within the backbone chain are least hydrated of all, although no physical or steric impediment seems to exist that would deny access to these oxygen atoms by water molecules.

Dietary cysteine regulates the levels of mRNAs encoding a family of cysteine-rich proteins of wool.

The abomasal or intravenous infusion of sulphur-containing amino acids such as cysteine or methionine into sheep on low-quality diets increases the sulphur content of the wool by increasing the synthesis of proteins containing a cysteine content of approximately 30 mol %. To investigate the molecular and cellular basis of this nutritional effect, quantitative analyses of wool keratin mRNA and protein levels, and follicle cortical cell type, were undertaken in sheep intravenously infused with cysteine. Northern blot analyses revealed that the mRNA levels of one gene family encoding cysteine-rich keratin-associated proteins (KAP4 family) expressed in the wool follicle cortex, increased approximately 5-6 times. Furthermore, the response was rapid as the mRNA levels increased approximately 3.5 times after 1 d of the cysteine infusion and, by 1 d post-infusion, they had fallen, approaching their basal level. No changes in the mRNA levels encoding the intermediate filament or the other keratin-associated protein families of lower cysteine content were observed. Concomitantly, two-dimensional polyacrylamide gel electrophoresis analysis of wool proteins showed a striking increase in the abundance of a group of cysteine-rich keratin-associated proteins in the wool by the end of the infusion period, returning to basal levels by 3 weeks later. At the cellular level, KAP4 expression was localized to the follicle paracortical cells, and the proportion of paracortical cells and the extent of KAP4 expression paralleled the changes in the cysteine infusion status of the sheep.

Excess thymidine induces folate sensitive fragile sites.

Folate sensitive fragile sites on human chromosomes have been found to be inducible in cultured lymphocytes by high levels of thymidine but not by high levels of BrdU. The biochemical interpretation of events leading to fragile site expression has been revised since it is now clear that low levels of either thymidylate or deoxycytidine triphosphate will result in this phenomenon. A model for the DNA at a fragile site, composed of alternating repeating polypurine/polypyrimidine sequences is proposed.

Translocation breakpoint in t(11;14) in B-cell leukemia is not at the rare fragile site at 11q13.3.

The cloned breakpoint at 11q13.3 of the t(11;14)(q13.3;q32.3) in a B-cell lymphocytic leukemia (B-CLL) was used to analyze DNA from individuals with and without the rare folate-sensitive fragile site at 11q13.3. On Southern blots there were no discernible differences. Subclones of the ends of the leukemia breakpoint clone were prepared and used for in situ hybridization to chromosomes expressing fra(11)(q13.3). Both subclones hybridized distal to the fragile site. These experiments indicate that the breakpoints at 11q13.3 in B-CLL (and in a B-cell lymphoma) are not at the fragile site at 11q13.3.

Sequence, expression, and evolutionary conservation of a gene encoding a glycine/tyrosine-rich keratin-associated protein of hair.

In hair differentiation several families of keratin proteins with distinctive amino acid compositions are produced. To study the role and regulation of one of these families, the glycine/tyrosine-rich keratin-associated proteins encoded by the KAP6 gene family, a partial wool follicle cDNA clone encoding a sheep KAP6 protein was sequenced and the corresponding gene isolated from a sheep cosmid library. The KAP6.1 gene encodes a basic protein of 82 amino acids (M(r) = 8,296) with a combined glycine and tyrosine content of approximately 60 mol%. There are several KAP6 genes in the sheep genome, all located within a 1,050-kilobase SfiI fragment. Northern blot analysis demonstrated that at least one member of the KAP6 family is expressed in the wool follicle. A rabbit KAP6 gene was isolated and its sequence and expression patterns were compared with the sheep gene. The sheep and rabbit genes have a nucleotide sequence identity of 89%, suggesting that they are equivalent genes and indicating strong selection pressure during evolution. Both genes contain several conserved sequence motifs of 7-9 nucleotides in their 5'-flanking regions that may be involved in the regulation of their expression. Localization of KAP6 mRNAs in sheep wool and rabbit hair follicles by in situ hybridization suggests that the genes are expressed in the cells of the hair shaft cortex in varying expression patterns. KAP6 expression starts relatively late in hair follicle differentiation, and the proportion of hair cortical cells that express it may change from follicle to follicle.

The crystal structure of auramine perchlorate: correlation of the results with specular reflection spectroscopy.

The x-ray analysis of auramine perchlorate has been performed in order to establish the orientation of the cationic chromophoric group in the crystal for comparison with the orientation that was proposed on the basis of specular reflection studies. The compound crystallizes with two molecules per unit cell in space group P1 and cell parameters a = 8.86 A, b = 9.69 A, c = 11.27 A, alpha = 97 degrees 40', beta = 99 degrees 34', and gamma = 106 degrees 16'. The orientation of the chromophore is in good agreement with the crystal model. Other features of the structure are the hydrogen bonding scheme of the perchlorate ion, the highly distorted chromophoric group, and the planar dimethylamine substituents.

Reversible bending and helix geometry in a B-DNA dodecamer: CGCGAATTBrCGCG.

A double-helical B-DNA dodecamer has been analyzed by single crystal x-ray diffraction methods and refined independently in four variants: sequence CGCGAATTCGCG at 20 degrees C and at 16 K, and CGCGAATTBrCGCG in 60% methylpentanediol at 20 and at 7 degrees C. The first three forms show a 14-19 degrees bend in overall helix axis, but the fourth is straight and unbent. Detailed comparisons of the various forms have led to a better understanding of helix geometry and bending. Structural principles can be understood best if organized under four headings: 1) intrinsic geometry of the sugar rings, 2) stacking and relative motion of base pairs, 3) geometry of the connecting phosphate backbone, and 4) mechanics of bending in B-DNA. The observed bending is neither completely localized nor smooth and continuous, but an intermediate compromise that can be termed "annealed kinking."

A linkage group with FRA16B (the fragile site at 16q22.1).

Polymorphic DNA markers located in bands 16q13, 16q21 and 16q22 were examined for recombination with FRA16B, the fragile site at 16q22.100. A tight linkage cluster D16S10-FRA16B-D16S4-HP was established. There were no recombinants (theta = 0.0, z = 8.3) between D16S10 and D16S4, which flank FRA16B. The markers D16S10 and D16S4 are in close proximity on the genetic map and delineate a small chromosomal segment, which contains the distamycin A-inducible fragile site.

Molecular genetics of human chromosome 16.

The major diseases mapped to chromosome 16 are adult polycystic kidney disease and those resulting from mutations in the alpha globin complex. There are at least six other less important genetic diseases which map to this chromosome. The adenine phosphoribosyltransferase gene allows for selection of chromosome 16 in somatic cell hybrids and a hybrid panel is available which segments the chromosome into six regions to facilitate gene mapping. Genes which have been mapped to this chromosome or which have had their location redefined since HGM8 include APRT, TAT, MT, HBA, PKD1, CTRB, PGP, HAGH, HP, PKCB, and at least 19 cloned DNA sequences. There are RFLPs at 13 loci which have been regionally mapped and can be used for linkage studies.

Fine mapping of gene probes and anonymous DNA fragments to the long arm of chromosome 16.

The fragile site, FRA16B, at 16q22.100 and four different translocations with breakpoints at 16q22.102, 16q22.105, 16q22.108, and 16q22.3 were used to locate and order DNA probes. This was achieved by Southern analysis of a somatic cell hybrid panel containing portions of chromosome 16 and by in situ hybridization. The anonymous DNA fragments D16S6, D16S10, and D16S11 were proximal to FRA16B and located at 16q13----q22.100. D16S4 and LCAT were located at 16q22.100----q22.102. TAT and HP were located at 16q22.105----q22.108. CTRB was located distal to 16q22.105 and therefore is in the distal half of 16q22. The order of markers in this region was determined as centromere-D16S6, D16S11, D16S10, MT-FRA16B-D16S4, LCAT-HP,TAT,CTRB-APRT- telomere. Linkage studies to determine map distances between the closest markers flanking the fragile site are now in progress.

A new location for the human adenine phosphoribosyltransferase gene (APRT) distal to the haptoglobin (HP) and fra(16)(q23)(FRA16D) loci.

The human adenine phosphoribosyltransferase gene (APRT) was mapped with respect to the haptoglobin gene (HP) and the fragile site at 16q23.2 (FRA16D). A subclone of APRT and a cDNA clone of HP were used for molecular hybridization to DNA from mouse-human hybrid cell lines containing specific chromosome 16 translocations. The APRT subclone was used for in situ hybridization to chromosomes expressing FRA16D. APRT was found to be distal to HP and FRA16D and was localized at 16q24, making the gene order cen-FRA16B-HP-FRA16D-APRT-qter.

Mapping the short arm of human chromosome 16.

Physical mapping of 13 different breakpoints on the short arm of chromosome 16 using previously mapped probes and the subsequent mapping of additional probes enabled the division of this portion of the chromosome into six different intervals. D16S94 was mapped between HBA and D16S80 and is closer to PKD1 than either HBA or D16S80. A tight linkage group which includes FRA16A, D16S8, and D16S79 was identified. Seven breakpoints, including FRA16A, could not be separated by probe localizations. This study provides the basis for the development of detailed maps of the short arm of chromosome 16.