Development of anti-Jk3 associated with silenced Kidd antigen expression and a novel single nucleotide variant of the JK gene.

Anti-Jk3 is a rare alloantibody to a high-prevalence antigen primarily seen in individuals of Polynesian descent and is associated with a handful of well-established variants of the SLC14A1 gene. We report a case of the Jknull phenotype, associated with formation of anti-Jk3, in a patient of non-Polynesian descent. This patient, a 51-year-old woman self-described as of Jamaican and Scottish ancestry, presented to our hospital for oncologic care. The patient's blood sample typed as blood group A, D+. All screening and panel reagent red blood cells showed reactivity, ranging from 2 to 4+; autocontrol and direct antiglobulin test were both negative. Antigen phenotyping revealed Jk(a-b-), leading to suspicion for anti-Jk3, which was subsequently confirmed by our immunohematology reference laboratory. Given her reported familial background, testing of the SLC14A1 gene was performed, revealing that the patient was heterozygous for the single nucleotide variant (SNV) at c.838G>A in exon 8 and therefore carries both JK*01 and JK*02 alleles that encode Jka and Jkb, respectively. However, the patient was found to be heterozygous for several additional SNVs: c.28G>A in exon 3; c.191G>A, c.226G>A, and c.303G>A in exon 4; and c.757T>C in exon 7. The patient's Jk(b-) phenotype can be explained by coinheritance of c.838A with c.191G>A, which defines null allele JK*02N.09. Coinheritance of SNVs c.28G>A and c.838G with rare SNV c.757C that is predicted to cause a non-conservative amino acid change (p.S253P) likely accounts for the complete serologic absence of Jka and the ability to form anti-Jk3 in this case. This finding would represent a new JK*01 null allele. This evaluation illustrates the importance of genetic analysis in identifying the factors preventing a high-prevalence antigen from being expressed, particularly when discovered outside of an expected racial or ethnic group.Anti-Jk3 is a rare alloantibody to a high-prevalence antigen primarily seen in individuals of Polynesian descent and is associated with a handful of well-established variants of the SLC14A1 gene. We report a case of the Jknull phenotype, associated with formation of anti-Jk3, in a patient of non-Polynesian descent. This patient, a 51-year-old woman self-described as of Jamaican and Scottish ancestry, presented to our hospital for oncologic care. The patient's blood sample typed as blood group A, D+. All screening and panel reagent red blood cells showed reactivity, ranging from 2 to 4+; autocontrol and direct antiglobulin test were both negative. Antigen phenotyping revealed Jk(a­b­), leading to suspicion for anti-Jk3, which was subsequently confirmed by our immunohematology reference laboratory. Given her reported familial background, testing of the SLC14A1 gene was performed, revealing that the patient was heterozygous for the single nucleotide variant (SNV) at c.838G>A in exon 8 and therefore carries both JK*01 and JK*02 alleles that encode Jka and Jkb, respectively. However, the patient was found to be heterozygous for several additional SNVs: c.28G>A in exon 3; c.191G>A, c.226G>A, and c.303G>A in exon 4; and c.757T>C in exon 7. The patient's Jk(b­) phenotype can be explained by coinheritance of c.838A with c.191G>A, which defines null allele JK*02N.09. Coinheritance of SNVs c.28G>A and c.838G with rare SNV c.757C that is predicted to cause a non-conservative amino acid change (p.S253P) likely accounts for the complete serologic absence of Jka and the ability to form anti-Jk3 in this case. This finding would represent a new JK*01 null allele. This evaluation illustrates the importance of genetic analysis in identifying the factors preventing a high-prevalence antigen from being expressed, particularly when discovered outside of an expected racial or ethnic group.

[Tapering of antidementia drugs, antidepressants and antipsychotics in elderly patients : When possible, when not?] / Antidementiva, Antidepressiva und Neuroleptika bei alten Patienten absetzen : Wann möglich, wann nicht?

BACKGROUND: Antidementia drugs, antidepressants and antipsychotics are among the most frequently prescribed medication in old and multimorbid patients. Due to side effects (e. g. prolonged QTc interval) in emergency medicine/intensive care unit or patients' wishes the question is often raised whether these drugs can be stopped and how this may be done. ANTIDEMENTIA DRUGS: If the cognition is stable under antidementia drugs or if the patient is in favour of the medication, it should be continued. After stopping antidementia drugs there may be a deterioration of cognitive function in the following 2-3 months. This should be discussed with the patient and the relatives/caregiver. ANTIDEPRESSANTS: In case of only slight or reactive depressive mood antidepressants should be tapered. The dose should be reduced over a period of at least 4 weeks. A sudden stop may cause a withdrawal syndrome with flu-like symptoms, fatigue, tremor, insomnia, anxiety or confusion. In severe depressive episodes there is a high risk of relapse; therefore deprescribing should only be done after a stable remission of 4-9 months. ANTIPSYCHOTICS: Antipsychotics in dementia or nursing home patients as well as in cases of delirium should be tapered, whereby confusion may increase again. When antipsychotics were prescribed because of hallucinations or severe psychosis, they should not be reduced or only with great caution.

[Staff knowledge of radiation protection in endovascular surgery]. / Connaissances du personnel de chirurgie endovasculaire en radioprotection.

INTRODUCTION: Image intensification exposes the endovascular surgery staff to ionizing radiation. The aim of this study was to determine awareness of ionizing radiation risks among personnel working in the endovascular surgery environment and the availability of radiation protection clothes and to propose appropriate corrective measures. METHODS: This descriptive study was performed in the endovascular operating theatre equipped with a mobile image intensifier unit in La Rabta vascular department in September 2017. We visited the endovascular theatre to identify the availability of radiation protection clothes. We used a questionnaire to identify personnel knowledge about ionizing radiation. We established a global score of knowledge to classify our population. RESULTS: We identified 85 professionals exposed to ionizing radiation. Sixty-four of them (75%) responded to our questionnaire; 65% were male; median age was 34 years (range: 25-61). Endovascular theatre personnel were surgeons (35%), nurses (34%), qualified technicians (18%) and other department employees (13%). The mean global score of knowledge was 8.15/20 (2-18). This score increased significantly with grade and seniority (Kruskal-Wallis test). CONCLUSION: In the present study, the results indicate insufficiency knowledge about radiation exposure among the endovascular staff and in radioprotection tools availability. In order to minimize all unnecessary radiation, attempts should be made to increase vascular theatre staff knowledge about radiation protection. Safety culture is a referral method to reduce radiation exposure as low as possible.

Are even geriatric patients becoming older and older? : Changes in the age structure of geriatric inpatients in the course of two decades.

BACKGROUND: The life expectancy of the German population has steadily risen in the course of the past decades. As especially the oldest members of the population are treated in geriatric clinics, it would be of interest to investigate whether the increase in population age can also be found among geriatric inpatients. PATIENTS AND METHODS: The demographic data of inpatients of a geriatric clinic in Hannover in the years 1994, 2004 and 2014 were analyzed according to age, gender and classification as acute care or geriatric rehabilitation. RESULTS: The mean patient age rose by 6 years in the past two decades. This was the case for both men and women but the age of men (+7.5 years) rose more than that of women (+4.9 years). Whereas the patient average age increased, especially in the first decade (+3.9 years), this increase slowed down in the following decade (+1.7 years). The 80 to 89-year-old patients remained the biggest and steadily increasing group (in 1994: 41.1%, 2004: 46.9% and 2014: 51.3%). The greatest increase, however, was found for those aged 90 years and older (1994: 4.8%, 2004: 12.2% and 2014: 17.7%). CONCLUSION: The results confirm the professional experiences of many geriatricians in that they care for an increasingly aging clientele. Particularly very old male patients in geriatric clinics are increasing. All health professional groups involved will have to face this challenge.

[Alzheimer dementia: course and burden on caregivers : Data over 18 months from German participants of the GERAS study]. / Alzheimer-Demenz: Verlauf und Belastung der Pflegepersonen : 18-Monats-Daten der deutschen Teilnehmer der GERAS-Studie.

The GERAS study is an international observational study with dementia patients of the Alzheimer type (AD) and their caregivers in everyday care. The 18-month data recorded in Germany are presented. Disease progression, medical and psychosocial consequences for both patients and caregivers were recorded using commonly used tests in clinical care: the mini mental status examination (MMSE), Alzheimer's disease assessment scale (ADAS-Cog14), Alzheimer's disease cooperative study activities of daily living inventory (ADCS-ADL), neuropsychiatric inventory (NPI-12), resource utilization in dementia (RUD) and the Zarit burden interview (ZBI). Definition of AD severity level (MMSE): 21-26 mild (miAD), 15-20 moderate (moAD), <15 moderately severe to severe (m/sAD). For the 550 participants (mean age: 75.2 years, SD 7.6 years), miAD (41.5%), moAD (28.4%) and m/sAD (30.2%), the MMSE worsened: in miAD by -2.4 (CI -3.1/-1.7), in moAD by -3.9 (CI -5.0/-2.8) and in m/sAD by -2.5 (CI -3.5/-1.5) at 18 months and the ADAS-Cog14 by 6.2 (miAD-CI 4.6/7.8) and 7.1 points (moAD CI 3.9/10.3). Changes in overall ADCS-ADL amounted to -8.4 (CI -10.1/-6.2) for miAD, -12.9 (CI -15.3/-10.4) for moAD and -10.2 points (CI-12.8/-7.7) for m/sAD. Caregiver burden (NPI-12) rose in miAD by 1.2 points (CI -0.2/2.2), in moAD by 3.4 (CI 1.8/5.1) and in m/sAD by 1.5 points (CI 0.2/3.3). At study start, the total time required by caregivers (RUD) was 3.1 h/day (SD 5.4 h/day) for miAD, 6.6 (SD 7.5) for moAD and 12.7 (SD 9.3) for m/sAD. With 4.4 (SD 9.4) h/day, the increase after 18 months was highest in moAD. Caregiver burden (ZBI) increased most markedly in moAD with 7.2 (CI 4.2/9.7), 90.7% of the patients received antidementia drugs, while 26.6% received psychotropic medication.

[Influence of cognition on treatment outcome in geriatric patients: association between MMSE and gain in activities of daily living]. / Einfluss der Kognition auf die Behandlungsergebnisse in der Geriatrie : Zusammenhang von MMSE und Zugewinn in den Alltagsaktivitäten.

BACKGROUND: Cognitive impairment or dementia influence the results of geriatric treatment. The aim of the study was to quantify this influence. PATIENTS AND METHODS: Data of 2527 patients from the years 2006 to 2009 were analysed in order to quantify the influence of cognition measured with the Mini Mental Status Examination (MMSE) on the improvement of activities of daily living as reflected by the Functional Independence Measure (FIM). RESULTS: Impaired cognition is accompanied by a lower FIM score on admission and on discharge. But the improvement of the FIM of slightly cognitively impaired patients (MMSE 20-26) is the same as in patients without cognitive impairment (MMSE 27-30). Patients with a MMSE below 20 points have smaller improvements in their FIM score but nevertheless 40 % of the patients with a MMSE of 10-19 and still 30 % of the patients with a MMSE of 0-9 points show better improvements than the average of all patients. CONCLUSION: Patients with a MMSE below 20 should not generally be excluded from geriatric treatment, but individual factors should be considered.

Alpha-lipoic acid as a new treatment option for Alzheimer's disease--a 48 months follow-up analysis.

Oxidative stress and neuronal energy depletion are characteristic biochemical hallmarks of Alzheimer's disease (AD). It is therefore conceivable that pro-energetic and antioxidant drugs such as alpha-lipoic acid might delay the onset or slow down the progression of the disease. In a previous study, 600mg alpha-lipoic acid was given daily to nine patients with AD (receiving a standard treatment with choline-esterase inhibitors) in an open-label study over an observation period of 12 months. The treatment led to a stabilization of cognitive functions in the study group, demonstrated by constant scores in two neuropsychological tests (the mini mental state exam, MMSE and the Alzheimer's disease assessment score cognitive subscale, ADAScog). In this report, we have extended the analysis to 43 patients over an observation period of up to 48 months. In patients with mild dementia (ADAScog < 15), the disease progressed extremely slowly (ADAScog: +1.2 points/year, MMSE: -0.6 points/year), in patients with moderate dementia at approximately twice the rate. However, the progression appears dramatically lower than data reported for untreated patients or patients on choline-esterase inhibitors in the second year of long-term studies. Despite the fact that this study was not double-blinded, placebo-controlled and randomized, our data suggest that treatment with alpha-lipoic acid might be a successful 'neuroprotective' therapy option for AD. However, a state-of-the-art phase II trial is needed urgently.

Alpha-lipoic acid as a new treatment option for Alzheimer [corrected] type dementia.

Oxidative stress and energy depletion are characteristic biochemical hallmarks of Alzheimer's disease (AD), thus antioxidants with positive effects on glucose metabolism such as thioctic (alpha-lipoic) acid should exert positive effects in these patients. Therefore, 600 mg alpha-lipoic acid was given daily to nine patients with AD and related dementias (receiving a standard treatment with acetylcholinesterase inhibitors) in an open study over an observation period of, on avarage, 337+/-80 days. The treatment led to a stabilization of cognitive functions in the study group, demonstrated by constant scores in two neuropsychological tests (mini-mental state examination: MMSE and AD assessment scale, cognitive subscale: ADAScog). Despite the fact that this study was small and not randomized, this is the first indication that treatment with alpha-lipoic acid might be a successful 'neuroprotective' therapy option for AD and related dementias.

The lysosomal targeting and intracellular metabolism of trypanosome lytic factor by Trypanosoma brucei brucei.

Trypanosome lytic factor (TLF) provides innate protection for humans against infection by the animal pathogen Trypanosoma brucei brucei but not against the agent of human African sleeping sickness, Trypanosoma brucei rhodesiense. TLF exists in two forms, TLF-1 and TLF-2. Prior studies suggested that TLF-1 causes lysosomal disruption and subsequent cell death in T. b. brucei. Here we confirm the lysosomal targeting of TLF-1 by immunolocalization with the trypanosome lysosomal membrane protein p67, and by co-fractionation of radiolabelled TLF-1 with lysosomal enzymes. In addition, pulse-chase studies indicate that TLF-1 is not degraded within the lysosome as compared to the host protein transferrin. In TLF-1 treated cells, transferrin is degraded normally, indicating that lysosomal proteases remain active during the early phase of TLF-1 treatment but fail to degrade TLF-1. Following endocytosis a TLF lipoprotein appears to undergo disulfide bond reduction prior to entering the lysosome. Results presented here indicate that TLF-1 lipoproteins are targeted to the lysosome but are resistant to trypanosome lysosomal proteases.

Virtual image grasping in patients with Alzheimer-type dementia.

Patients with various neuropsychiatric disorders who confused virtual images with real objects have recently been described. When asked to take objects from an investigator's hand while looking in a mirror, these patients reached directly into the mirror for the object's image. To investigate whether the phenomenon occurs in patients with Alzheimer's disease, we studied 127 cases, of whom 67 were suspected of having Alzheimer-type dementia (DAT). The phenomenon in question was not observed in either of two control groups, but whereas 38 (57%) of the DAT patients responded correctly, 17 (25%) of them grasped at the mirror image. Characteristically, none of the eight patients who grasped into the mirror and underwent single photon emission computed tomography analysis showed symmetric activity. We also analyzed the behaviour and verbal utterances of the DAT patients in front of the mirror.

Large-scale analysis of the yeast genome by transposon tagging and gene disruption.

Economical methods by which gene function may be analysed on a genomic scale are relatively scarce. To fill this need, we have developed a transposon-tagging strategy for the genome-wide analysis of disruption phenotypes, gene expression and protein localization, and have applied this method to the large-scale analysis of gene function in the budding yeast Saccharomyces cerevisiae. Here we present the largest collection of defined yeast mutants ever generated within a single genetic background--a collection of over 11,000 strains, each carrying a transposon inserted within a region of the genome expressed during vegetative growth and/or sporulation. These insertions affect nearly 2,000 annotated genes, representing about one-third of the 6,200 predicted genes in the yeast genome. We have used this collection to determine disruption phenotypes for nearly 8,000 strains using 20 different growth conditions; the resulting data sets were clustered to identify groups of functionally related genes. We have also identified over 300 previously non-annotated open reading frames and analysed by indirect immunofluorescence over 1,300 transposon-tagged proteins. In total, our study encompasses over 260,000 data points, constituting the largest functional analysis of the yeast genome ever undertaken.

Design strategies and performance of custom DNA sequencing primers.

This study surveyed strategies of sequencing primer selection and evaluated primer performance in automated DNA sequencing. We asked participants to relate their preferred primer design strategies to identify primer characteristics that are considered most important in sequencing primer design. The participants preferred primers of 18-24 nucleotides (nt), 39%-58% G + C, a melting temperature (Tm) of 53 degrees-65 degrees C with a 1-2 nt 3' GC clamp, hairpin stems of less than 2-3 bp, homopolymeric runs of less than 4-5 nt, primer dimers of less than 3-4 bp and secondary priming sites of less than 3-4 bp. We provided a 300-bp test sequence and asked participants to submit sequences of 1-3 optimal sequencing primers. Submitted primers ranged from 17-24 nt and largely conformed to the preferred parameters. Submitted primers were distributed across the test sequence, although some sites were disfavored. Surprisingly, approximately 45% of the primers were selected "manually", more than by any software package. Each of 69 submitted and 95 control primers, distributed at 3-bp intervals across the test sequence, were synthesized, purified and tested using a Model 377 PRISM DNA Sequencer with dichlororhodamine dye terminator reagents (dRhodamine dye terminators). Approximately half of the control primers were also tested using rhodamine dye terminator reagents ("old" rhodamine dye terminators). The results indicated that primer physico-chemical characteristics thought to have a strong impact on sequencing performance had surprisingly little effect. Thus, primers with high or low percent G + C or Tm, strong secondary priming scores or long 3' homopolymeric stretches yielded excellent sequences with the dRhodamine dye terminator reagents, although these characteristics had a stronger effect when the old rhodamine reagents were used. The old rhodamine reagents gave sequences with a similar average read length, but the number of errors and ambiguities or "N's" was consistently higher. Moreover, the effects of the primer physico-chemical characteristics were also more evident with the old rhodamine dyes. We conclude that under optimal sequencing conditions with highly pure template and primer, many of the commonly applied primer design parameters are dispensable, particularly when using one of the new generation of sequencing reagents such as the dichlororhodamine dye terminators.

The nuclear envelope serves as an intermediary between the ER and Golgi complex in the intracellular parasite Toxoplasma gondii.

Morphological examination of the highly polarized protozoan parasite Toxoplasma gondii suggests that secretory traffic in this organism progresses from the endoplasmic reticulum to the Golgi apparatus using the nuclear envelope as an intermediate compartment. While the endoplasmic reticulum is predominantly located near the basal end of the parasite, the Golgi is invariably adjacent to the apical end of the nucleus, and the space between the Golgi and nuclear envelope is filled with numerous coatomer-coated vesicles. Staining with antiserum raised against recombinant T. gondii beta-COP confirms its association with the apical juxtanuclear region. Perturbation of protein secretion using brefeldin A, microtubule inhibitors or dithiothreitol disrupts the Golgi, causing swelling of the nuclear envelope, particularly at its basal end. Prolonged drug treatment leads to gross distention of the endoplasmic reticulum, filling the basal end of the parasite. Cloning and sequencing of the T. gondii homolog of the chaperonin protein BiP identifies the carboxy-terminal amino acid sequence HDEL as this organism's endoplasmic reticulum-retention signal. Appending the HDEL motif to a recombinant secretory protein (a chimera between the parasite's major surface protein fusion, P30, and the Green Fluorescent Protein) causes this secretory reporter to be retained intracellularly. P30-GFP-HDEL fluorescence was most intense within the nuclear envelope, particularly at the apical end. These data support a model of secretion in which protein traffic from the endoplasmic reticulum to Golgi occurs via the apical end of the nuclear envelope.

Use of transdermal amitriptyline gel in a patient with chronic pain and depression.

A man with severe inflammatory bowel disease suffered from chronic abdominal pain and depression. A transdermal amitriptyline gel preparation was compounded since he was unable to take drugs orally Serum concentrations of amitriptyline and its active metabolite nortriptyline were measured over 24 hours. Symptoms of depression were monitored before starting transdermal therapy and at the end of 6 weeks. Pain symptoms and amitriptyline adverse drug events were monitored daily Steady-state serum concentrations of drug and metabolite were within the therapeutic range over 24 hours. The patient reported that his mood was improved but his abdominal pain remained unchanged. Transdermal amitriptyline gel was well tolerated and is an alternative delivery system in patients unable to take drugs orally.

Transport and trafficking: Toxoplasma as a model for Plasmodium.

Like Plasmodium, the protozoan parasite Toxoplasma gondii is a member of the phylum Apicomplexa, and an obligate intracellular pathogen. Unlike Plasmodium, however, Toxoplasma is highly amenable to experimental manipulation in the laboratory. The development of molecular transformation protocols for T. gondii has provided both scientific precedent and practical selectable markers for Plasmodium. Beyond the feasibility of molecular biological experimentation now possible in both systems, the high frequency of stable transformation in Toxoplasma allows this parasite to be used for molecular genetic analysis. The ability to control homologous vs. non-homologous recombination in T. gondii permits gene knockouts/allelic replacements at previously cloned loci, and saturation insertional mutagenesis of the entire parasite genome (and cloning of the tagged loci). T. gondii also exhibits unusual ultrastructural clarity, facilitating cell biological analysis. The accessibility of Toxoplasma as an experimental system allows this parasite to be used as a surrogate for asking many questions that cannot easily be addressed in Plasmodium itself. T. gondii also serves as a model system for genetic exploration of parasite biology and host-parasite interactions. Success stories include: biochemical analysis of antifolate resistance mechanisms; pharmacological studies on the mechanisms of macrolide activity; genetic identification of nucleobase/nucleoside transporters and metabolic pathways; and cell biological characterization of the apicomplexan plastid. As with any model system, not all questions of interest to malariologists can be addressed in Toxoplasma; differentiating between sensible and foolish questions requires familiarity with the biological similarities and differences of these systems.