p.(Asp47Asn) and p.(Thr62Met): non deleterious LDL receptor missense variants functionally characterized in vitro.

Familial Hypercholesterolemia (FH) is a common genetic disorder caused most often by mutations in the Low Density Lipoprotein Receptor gene (LDLr) leading to high blood cholesterol levels, and ultimately to development of premature coronary heart disease. Genetic analysis and subsequent cascade screening in relatives allow diagnosis of FH at early stage, especially relevant to diagnose children. So far, more than 2300 LDLr variants have been described but only a minority of them have been functionally analysed to evaluate their pathogenicity in FH. Thus, identifying pathogenic mutations in LDLr is a long-standing challenge in the field. In this study, we investigated in vitro the activity p.(Asp47Asn) and p.(Thr62Met) LDLr variants, both in the LR1 region. We used CHO-ldlA7 transfected cells with plasmids carrying p.(Asp47Asn) or p.(Thr62Met) LDLr variants to analyse LDLr expression by FACS and immunoblotting, LDL binding and uptake was determined by FACS and analysis of mutation effects was assessed in silico. The in vitro activity assessment of p.(Asp47Asn) and p.(Thr62Met) LDLr variants shows a fully functional LDL binding and uptake activities. Therefore indicating that the three of them are non-pathogenic LDLr variants. These findings also emphasize the importance of in vitro functional LDLr activity studies to optimize the genetic diagnosis of FH avoiding the report of non-pathogenic variants and possible misdiagnose in relatives if cascade screening is carried out.

Replacement of cysteine at position 46 in the first cysteine-rich repeat of the LDL receptor impairs apolipoprotein recognition.

BACKGROUND AND AIMS: Pathogenic mutations in the Low Density Lipoprotein Receptor gene (LDLR) cause Familial Hypercholesterolemia (FH), one of the most common genetic disorders with a prevalence as high as 1 in 200 in some populations. FH is an autosomal dominant disorder of lipoprotein metabolism characterized by high blood cholesterol levels, deposits of cholesterol in peripheral tissues such as tendon xanthomas and accelerated atherosclerosis. To date, 2500 LDLR variants have been identified in the LDLR gene; however, only a minority of them have been experimentally characterized and proven to be pathogenic. Here we investigated the role of Cys46 located in the first repeat of the LDL receptor binding domain in recognition of apolipoproteins. METHODS: Activity of the p.(Cys46Gly) LDLR variant was assessed by immunoblotting and flow cytometry in CHO-ldlA7 expressing the receptor variant. Affinity of p.(Cys46Gly) for LDL and VLDL was determined by solid-phase immunoassays and in silico analysis was used to predict mutation effects. RESULTS AND CONCLUSION: Functional characterization of p.(Cys46Gly) LDLR variant showed impaired LDL and VLDL binding and uptake activity. Consistent with this, solid-phase immunoassays showed the p.(Cys46Gly) LDLR variant has decreased binding affinity for apolipoproteins. These results indicate the important role of Cys46 in LDL receptor activity and highlight the role of LR1 in LDLr activity modulation. This study reinforces the significance of in vitro functional characterization of LDL receptor activity in developing an accurate approach to FH genetic diagnosis. This is of particular importance because it enables clinicians to tailor personalized treatments for patients' mutation profile.

Hipnosis clínica para mejorar la disposición a la curación de un paciente en la unidad de cuidados intensivos / Clinical hypnosis to improve a patient's disposition to receive treatment in an ICU / Hipnose clínica para melhorar a disposição à cura de um paciente em UCI

En el presente artículo se revisa el caso de una persona adulta hospitalizada en una unidad de paciente crítico, quien ha evolucionado con agitación durante la estadía hospitalaria y que requiere curación de úlceras por presión. A la valoración la enfermera se anticipa a la reacción negativa que podría tener el paciente ante el dolor asociado a la técnica, por lo cual utiliza la hipnosis clínica con el objetivo de favorecer la disposición del paciente a mejorar su comodidad; para lograr realizar la curación sin signos de dolor en la monitorización ni en el reporte subjetivo, se constata el mejoramiento del bienestar general del paciente relacionado con la relajación involucrada en el proceso. Se presenta el proceso de enfermería, la descripción del procedimiento y se motiva a los profesionales de enfermería a incorporar la hipnosis dentro de su repertorio de competencias para la gestión del cuidado.

In this article, we review the case of an adult person hospitalized in a critical patient unit, who has become agitated, and who requires treatment for pressure sores. During assessment, the nurse anticipates the patient's possible negative reaction associated to the pain, and thus uses the clinical hypnosis technique in order to favor the patient's better disposition. This procedure improved the patient's comfort, and the healing treatment was performed without signs of pain. As a result of the enhanced relaxation, the patient's overall wellbeing was ensured. The nursing process is presented as well as the specific procedure. Nursing professionals are encouraged to incorporate the hypnosis into their competences repertoire for the care management.

No presente artigo revisa-se o caso de uma pessoa adulta hospitalizada em uma unidade de paciente crítico, quem tem evoluído com agitação durante a estadia hospitalar e que requer cura de úlceras por pressão. À avaliação, a enfermeira antecipa-se à reação negativa que poderia ter o paciente perante a dor associada à técnica, pelo qual utiliza a hipnose clínica com o objetivo de favorecer a disposição do paciente para melhorar seu conforto, consegue-se realizar a cura sem signos de dor na monitorizarão nem no relatório subjetivo, constata-se a melhora do bem-estar geral do paciente relacionado com a relaxação envolvida no processo. Apresenta-se o processo de enfermagem, a descrição do procedimento e motiva-se aos profissionais de enfermagem a incorporar a hipnose dentro de seu repertório de competências para a gestão do cuidado.

Middle and Later Stone Age large mammal and tortoise remains from Die Kelders Cave 1, Western Cape Province, South Africa.

Die Kelders Cave 1, South Africa, has provided more than 150,000 taxonomically identifiable mammal and tortoise bones from Middle Stone Age (MSA) and Later Stone Age (LSA) deposits. Cape dune mole-rats dominate the mammal sample, and they appear to have been accumulated mainly by people during the LSA occupation and mainly by eagle owls in the MSA. In sharp contrast to the LSA fauna, the MSA sample contains extralimital ungulates that imply relatively moist, grassy conditions. The large mean size of the MSA mole-rats also points to greater humidity, while the large size of the gray mongooses implies cooler temperatures. The sum supports luminescence and ESR dates that place the MSA occupation within the early part of the Last Glaciation (global isotope stage 4). The Die Kelders ungulate bones support those from Klasies River Mouth in suggesting that MSA people obtained dangerous terrestrial prey much less frequently than their LSA successors, probably because MSA people lacked the bow and arrow and other projectile weapons. The Die Kelders tortoise bones constrain the extent of climatic change, since their abundance indicates that warm, dry days remained common, at least seasonally. The tortoises tend to be much larger in the MSA layers than in the LSA ones, suggesting that MSA people collected tortoises less intensively, probably because MSA populations were relatively sparse.

Duinefontein 2: an Acheulean site in the Western Cape Province of South Africa.

Excavations at Duinefontein (DFT) 2 near Cape Town, South Africa have recovered numerous stone artefacts and animal bones on an ancient surface sealed within iron-stained eolian sands. U-series analysis of an overlying calcrete places the sands before 150 ka ago, while the large mammal taxa imply an age between 400 and 200 ka ago. The artefacts include a classic Acheulean handaxe and probable biface shaping flakes that support this age estimate. The principal mammalian species are long-horned buffalo, black wildebeest, greater kudu, Cape zebra, and grysbok/steenbok, which imply a grass-and-bush mosaic instead of the historic small-leafed shrubland. Hippopotamus and reedbuck indicate that water stood nearby, probably in dune swales. The large mammal bones are mostly vertebrae and other axial elements, often in near-anatomical order. Both proximal and distal appendicular elements are rare. Bones with carnivore damage are common, but ones with stone tool marks are scarce. The sum suggests a water-edge attritional death site where people played a minimal role and carcasses were disarticulated mainly by carnivore feeding and by trampling. Stone tool marks tend to be equally rare at other Acheulean attritional death sites, and the implication may be that Acheulean people rarely obtained large mammals, whether by hunting or scavenging. Human scavengers at DFT2 would not have encountered a disproportionate number of distal (versus proximal) limb elements, and it follows that the tendency for distal elements to dominate many archeological assemblages need not reflect scavenging versus hunting. Even if DFT2 was not itself a locus of intense human activity, it provides a useful baseline for evaluating bone damage, skeletal part representation, and other variables at sites where people were deeply involved.