Tackle your Tics, a brief intensive group-based exposure treatment for young people with tics: results of a randomised controlled trial.

Tics can have a serious impact on the quality of life of children and their families. Behavioural therapy is an evidence-based first line treatment for tic disorders. This randomised controlled trial studied the efficacy of a brief, condensed group-based programme for children with tics (Dutch Trial Registry NL8052, 27 September 2019). Tackle your Tics is a four-day group treatment, including exposure and response prevention and supporting components, delivered by therapists and 'experts by experience'. We collected outcome measures at baseline (T1), directly post-treatment (T2), and at three- and 6-months follow-up (T3, T4) including tic severity (primary outcome measure), tic-related impairment, quality of life, tic-related cognitions, emotional/behavioural functioning, family functioning, treatment satisfaction and adherence. Outcomes directly post-treatment improved in both the treatment group (n = 52) and waiting list (n = 54), but showed no statistically significant differences between the conditions (differential change over time T1-T2) on tic severity (Yale Global Tic Severity Scale), quality of life (Gilles de la Tourette Syndrome Quality of Life Scale), tic-related cognitions and family functioning. At longer term (T3), again no between-group difference was found on tic severity, but tic-related impairment, quality of life and emotional/behavioural functioning significantly improved in the treatment group compared to the waiting list. Mean treatment satisfaction scores were favourable for both children and parents. Directly posttreatment, Tackle your Tics showed no superior effect compared to waiting list. However, on longer term this brief four-day group treatment was effective in improving tic-related impairment, quality of life and emotional/behavioural functioning.

Tackle your Tics: pilot findings of a brief, intensive group-based exposure therapy program for children with tic disorders.

Tourette syndrome (TS) and other chronic tic disorders (CTD) are prevalent neurodevelopmental disorders, which can have a huge burden on families and society. Behavioral treatment is a first-line intervention for tic disorders. Despite demonstrated efficacy, tic reduction and utilization rates of behavioral treatment remain relatively low. Patient associations point to an urgent need for easy-to-undergo treatments that focus both on tic reduction and improvement of quality of life. To enhance treatment outcome and overcome treatment barriers, this pilot study's aim was to investigate the feasibility and preliminary results of a brief, intensive group-based treatment. Tackle your Tics is a 4-day intensive and comprehensive group-based program for children and adolescents (9-17 years) with a tic disorder, consisting of exposure and response prevention (ERP) treatment and additional supporting components, such as coping strategies, relaxing activities and parent support. Assessments were performed pre- and post-treatment and at 2 months follow-up, to test outcomes on tic severity and quality of life, and explore premonitory urges, emotional and behavioral functioning and treatment satisfaction (N = 14, of whom 13 completed the treatment). Parents and children rated this treatment positive on a treatment satisfaction questionnaire. On tic severity (Yale Global Tic Severity Scale) and quality of life (Gilles de la Tourette Syndrome Quality of Life Scale for children and adolescents), improvements between pre-treatment and follow-up were found. Intensive ERP in group format is promising as a feasible treatment to improve both tic severity as well as quality of life. Larger controlled trials are needed to establish its effectiveness.

Oocyte insemination with the Walking Egg simplified IVF culture system - an investigation into reduced sperm numbers, sperm DNA fragmentation and reactive oxygen species formation.

RESEARCH QUESTION: What is the lowest number of sperm that can be used for oocyte insemination during either conventional or the Walking Egg simplified IVF? Does the use of low numbers of sperm in high volume (1 ml) culture media have an effect on sperm DNA fragmentation and reactive oxygen species formation? Also, does the extended co-incubation of embryos with sperm and cumulus cells in the Walking Egg culture tubes induce higher levels of reactive oxygen species? DESIGN: Binding of sperm to the zona pellucida was compared using a modified hemi-zona assay. In the first part of the study, the binding capacity of decreasing concentrations of motile spermatozoa was evaluated, followed by a comparison of sperm binding after simulated insemination by conventional or the Walking Egg simplified culture protocol. Sperm DNA fragmentation was determined between test and control samples in the second part of the study and reactive oxygen species was measured in spent culture media. As a supplementary examination, reactive oxygen species formation, with the simulated co-incubation of cumulus and sperm cells, was compared between the conventional and Walking Egg IVF culture systems. RESULTS: Sperm-zona binding in 50 µl culture media, indicated mean sperm binding of more than 20 sperm per hemi-zona with as low as 1000 sperm used for insemination. Using a higher volume of culture media, as is done in the Walking Egg simplified IVF culture system, resulted in 42.8% reduced sperm-zona binding. No significant difference in DNA integrity was observed between the two test groups. The amount of ROS generated during conventional IVF in the first 18 hours of incubation was more than that produced in the simplified culture system over sixty-six hours. Only during extended culture for 114 hours in the simplified culture system, did the ROS generated slightly surpass that of conventional IVF at 18 hours. CONCLUSION: Oocyte insemination with as little as 2 x 103 motile sperm showed sufficient sperm-zona binding capacity to be indicative of fertilization potential, supporting the Walking Egg simplified IVF insemination protocol. No difference in DNA fragmentation was observed between conventional and the simplified IVF culture systems, while reactive oxygen species formation was indicated to be at a slower rate during incubation with the Walking Egg simplified IVF culture system than with conventional IVF.

Mitochondrial transfer: Implications for assisted reproductive technologies.

The use of mitochondrial transfer as a clinic procedure is drawing closer to reality. Here we provide a detailed overview of mitochondrial transfer techniques - both established and recent - including pronuclear, spindle, ooplasmic and blastomere transfer. Reasons as to why some techniques are more suitable for the prevention of mitochondrial DNA disease than others, as well as the advantages and disadvantages of each methodology, are discussed. The possible clinical introduction of these techniques has raised concerns about the adverse effects they may have on resultant embryos and offspring. Success rates of each technique, embryo viability and developmental consequences post mitochondrial transfer are addressed through analysis of evidence obtained from both animal and human studies. Counterarguments against potential mitochondrial-nuclear genome incompatibility are also provided. Additional clinical applications of mitochondrial transfer techniques are discussed. These include the rescue or enhancement of fertility in women of advanced maternal age or those suffering from diabetes. An alternative to using mitochondrial DNA transfer for germ line therapies is the therapeutic use of somatic cell nuclear transfer for the generation of personalised stem cells. Although ethically challenging, this method could offer patients already suffering from mitochondrial DNA diseases a novel treatment option.

Large volume cryoprotectant-free vitrification: an alternative to conventional cryopreservation for human spermatozoa.

Vitrification is a simple and cost-effective method for the storage of human spermatozoa without the use of conventional cryoprotectants, by plunging the sperm suspension directly into liquid nitrogen. As a result, solidification of living cells without the formation of ice crystals is achieved during cooling. This study aimed to compare cryoprotectant-free vitrification to conventional cryopreservation protocols. Semen samples (n = 35) were collected from patients seeking diagnostic assistance at the Reproductive and Endocrine Unit at Steve Biko Academic Hospital. Samples were processed using a discontinuous density-gradient centrifugation method. Washed samples were split into two aliquots and cryopreserved either by means of cryoprotectant-free vitrification (sucrose + 1% albumin) or conventional slow freezing (TEST-yolk buffer). Post-thawing, the sperm motion parameters, mitochondrial membrane potential (Δψm) and DNA fragmentation were compared between the two groups. No significant differences were observed in the sperm motility parameters (P > 0.05). Significantly higher percentages of Δψm (11.99% ± 4.326% versus 6.58% ± 1.026%; P < 0.001) and lower percentages of DNA fragmentation (2.79% ± 1.017% versus 3.86% ± 1.38%; P < 0.01) were observed when comparing cryoprotectant-free vitrification to conventional cryopreservation. Cryoprotectant-free vitrification is a rapid and promising alternative to conventional methods resulting in good-quality spermatozoa post-thaw.

Prevention of infections in an ART laboratory: a reflection on simplistic methods.

Preventative measures combined with reactive remedial actions are generic management tools to optimize and protect an entity's core businesses. Differences between assisted reproduction technology (ART) laboratories in developing versus developed countries include restricted access to, or availability of resources, and the prevalence of pathological conditions that are endemic or common in non-industrialized regions. The aim of this paper is to discuss the prevention of infections in an ART laboratory in a low to middle-income country, with reference to simplistic risk reduction applications to avoid the introduction and transmission of pathogens. Diagnostic and procedural phases will be examined, i.e. (i) screening for microbes during patient evaluation, and (ii-iii) prevention of environmental and procedural contamination. Preventative action is enabled by knowledge of threats and the degree of risk involved. Awareness and understanding of the vulnerabilities in an ART system, wherein laboratory personnel operate, are invaluable assets when unforeseen equipment failure occurs or instant decisions have to be made to safeguard procedures. An inter-connective team approach to patient treatment, biosafety training and utilization of practical procedures such as semen decontamination, are fundamental tools in a laboratory's risk-reduction armoury to prevent and eliminate infectious elements.

Psychiatric symptoms causing delay in diagnosing childhood cancer: two case reports and literature review.

INTRODUCTION: A somatic disorder may initially be overlooked when a child presents with psychiatric symptoms. We report two children with anorexia nervosa as initial diagnosis and in whom there was a delay in the final diagnosis of the underlying malignancy. A literature survey was performed including patients under 18 years of age with psychiatric symptoms in whom later on an oncological diagnosis became evident as an explanation. RESULTS: We have found 30 additional cases, with a median delay of 12 months until the diagnosis of the tumour. Overall, 16 boys and 16 girls had a solid tumour: 26 central nervous system tumours, 3 tumours of the gastrointestinal tract and 3 others. In 25 out of 32 patients anorexia nervosa was assumed, although it always appeared to be atypical. Patients younger than 7 years had a significantly longer delay until final diagnosis, while no other patient characteristics correlated with such delay. DISCUSSION: In addition to careful physical (including full neurological) examination, we advise additional neuroimaging especially in each case of atypical presentation of anorexia nervosa, in order to avoid a delay in diagnosis of a possible malignancy. Furthermore, it is desirable to perform a re-examination when a psychiatric disorder does not respond to therapy, in order not to overlook an underlying oncological disease.

ART in South Africa: The price to pay.

In developing countries especially in Sub-Saharan Africa, human immunodeficiency virus (HIV) infection -together with limited resources adds to the hindrances in becoming a parent. Although the South African's Bill of Rights proclaim that South Africans can "make decisions concerning reproduction"; access to and the use of Assisted Reproduction Technology (ART) are viewed in general as excessively expensive, accessible only to the privileged few. A dissection of cost-drivers within an ART laboratory, such as procedures; sperm preparations; laboratory supplies including embryo culture media and cryopreservation are discussed in the current overview. Subject to the nature of an ART practice, i.e. private vs. public/tertiary, the structure of a unit will vary with regards to patient demographics, costs and services offered. The average fees per procedure for 20 practices in the private sector in South Africa are: (i) IUI: €â€ˆ542 ± €159, (ii) IVF: €â€ˆ3,255 ± €â€ˆ576 and (iii) ICSI: €3,302 ± €â€ˆ625. Laboratory costs can contribute between 35 and 48% of ART fees payable in the private sector. Low-cost public ART services are available to citizens of the country at a few tertiary academic units. Some private practices also cater specifically for middle-income citizens. ART procedures need not be propelled towards the must-have and cannot-do without approach, but providers should also reflect on the validity of the techniques and equipment, without compromising treatment virtue.

Treatment of human sperm with serine protease during density gradient centrifugation.

PURPOSE: Seminal pathogens can bind specifically or non-specifically to spermatozoa, rendering semen decontamination procedures ineffective, whereby vertical or horizontal transmission of the infection could occur. Serine proteases have been demonstrated to effectively inactivate viruses and to break pathogen-sperm bonds. However, the addition of a protease to density gradient layers during semen processing could negatively impact on sperm parameters. This study investigated the effect of the addition of a recombinant, human-sequence protease (rhProtease) on sperm parameters during density gradient centrifugation. METHODS: (i) Pooled semen samples (n = 9) were split and processed by density gradient centrifugation, with the top density layers supplemented, or non-supplemented with rhProtease at three different concentrations (diluted 2, 10 and 20 times). Sperm parameters were then analysed by flow cytometry and computer-assisted semen analyses. (ii) Semen samples (n = 5) were split and similarly processed using PureSperm® Pro, with rhProtease in the 40 % density gradient layer, or standard PureSperm® not supplemented with rhProtease (Nidacon, International) respectively. The Hemizona assay was then utilized to compare sperm-zona binding post processing. RESULTS: Evaluation of sperm parameters indicated that rhProtease did not, at any of the tested concentrations, have an impact on (i) mitochondrial membrane potential, vitality, motility, or (ii) zona binding potential. CONCLUSION: We report that the addition of rhProtease to density gradients is a non-detrimental approach that could improve the effectiveness of semen processing for the elimination of seminal pathogens, and benefit assisted reproduction outcome.

Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel tube insert.

The occurrence of bacteria in sperm samples intended for in vitro fertilisation can compromise the outcome of assisted reproductive techniques. Effective semen processing procedures should therefore be implemented to remove bacteria from semen. Unfortunately, technique failure does occur whereby bacteria can be found in processed sperm preparations. To improve the effectiveness of semen processing, a novel centrifuge tube insert was developed to facilitate the layering of density gradients and semen, and to prohibit the re-infection of purified sperm pellets. The purpose of this study was to: (i) determine the prevalence and type of bacteria present in semen of patients participating in the Unit's assisted reproduction program and (ii) evaluate the effectiveness of density gradient centrifugation with the novel tube insert, for the elimination of bacteria and yeast from spiked human semen samples. A survey in 2007-2010 indicated that 50% of semen samples were found to have positive bacterial cultures. Semen processing by means of density gradient centrifugation with the novel tube insert eliminated significantly more in vitro derived (spiked) bacteria and yeast from semen compared to processing without the insert (P < 0.004). Therefore, it is highly recommended that the centrifuge tube insert, ProInsert™, be incorporated into assisted reproductive programs.

The effect of pulsed 900-MHz GSM mobile phone radiation on the acrosome reaction, head morphometry and zona binding of human spermatozoa.

Several recent studies have indicated that radiofrequency electromagnetic fields (RF-EMF) have an adverse effect on human sperm quality, which could translate into an effect on fertilization potential. This study evaluated the effect of RF-EMF on sperm-specific characteristics to assess the fertilizing competence of sperm. Highly motile human spermatozoa were exposed for 1 h to 900-MHz mobile phone radiation at a specific absorption rate of 2.0 W/kg and examined at various times after exposure. The acrosome reaction was evaluated using flow cytometry. The radiation did not affect sperm propensity for the acrosome reaction. Morphometric parameters were assessed using computer-assisted sperm analysis. Significant reduction in sperm head area (9.2 ± 0.7 µm² vs. 18.8 ± 1.4 µm²) and acrosome percentage of the head area (21.5 ± 4% vs. 35.5 ± 11.4%) was reported among exposed sperm compared with unexposed controls. Sperm-zona binding was assessed directly after exposure using the hemizona assay. The mean number of zona-bound sperm of the test hemizona and controls was 22.8 ± 12.4 and 31.8 ± 12.8 (p < 0.05), respectively. This study concludes that although RF-EMF exposure did not adversely affect the acrosome reaction, it had a significant effect on sperm morphometry. In addition, a significant decrease in sperm binding to the hemizona was observed. These results could indicate a significant effect of RF-EMF on sperm fertilization potential.

Effects of Non-Thermal Mobile Phone Radiation on Breast Adenocarcinoma Cells

Mobile phone usage currently exceeds landline communication in Africa. The extent of this usage has raised concerns about the long-term health effects of the ongoing use of mobile phones. To assess the physiological effects of radiation from mobile phones in vitro; MCF-7 breast adenocarcinoma cells were exposed to 2W/kg non-thermal 900-MHz mobile phone radiation. The effects investigated were those on metabolic activity; cell morphology; cell cycle progression; phosphatidylserine (PS) externalisation and the generation of reactive oxygen species and nitrogen species. Statistically insignificant increases in mitochondrial dehydrogenase activity were observed in irradiated cells when compared to controls. Fluorescent detection of F-actin demonstrated an increase in F-actin stress fibre formation in irradiated MCF-7 cells. Cell cycle progression revealed no statistically significant variation. A small increase in early and late apoptotic events in irradiated MCF-7 cells was observed. No statistically significant changes were observed in reactive oxygen and reactive nitrogen species generation. In addition; quantitative and qualitative analyses of cell cycle activity and nuclear and cytosolic changes; respectively; revealed no significant changes. In conclusion; exposure to 1 h of 900-MHz irradiation induced an increase in PS externalisation and an increase in the formation of F-actin stress fibres in MCF-7 cells. Data obtained from this study; and their correlation with other studies; provides intriguing links between radio frequency radiation and cellular events and warrant further investigation

[Olanzapine in the treatment of adolescents with anorexia nervosa]. / Olanzapine in de behandeling van adolescenten met anorexia nervosa.

This article, which was triggered by a case study of a 15-year-old female patient, gives an overview of the literature on the use of olanzapine as an adjunctive treatment for anorexia nervosa in adolescents. On the basis of studies performed so far (two small double-blind placebo-controlled studies, two open-label trials, one retrospective study, a number of case studies that included adolescents, and four series of case studies on adolescents alone), the short-term results of using olanzapine were promising. However, careful monitoring is needed.

Comparison between propidium iodide and 7-amino-actinomycin-D for viability assessment during flow cytometric analyses of the human sperm acrosome.

Evaluation of the acrosome reaction can shed light on the fertilising competence of spermatozoa. To eliminate false-positive results when evaluating the acrosome status of human sperm cells, two viability probes propidium iodide (PI) and 7-amino-actinomycin D (7-AAD) were compared for their ability to stain nonviable cells post-fixation and permeabilisation. Both the mean fluorescence and % dead cells differed significantly with time (P < 0.0001). Unlike PI, 7-AAD did not leach from cells and fluorescence remained stable for up to 4 h. Furthermore, 7-AAD proved to be a proficient marker to exclude dead sperm cells during flow cytometric evaluation of ionophore-induced acrosome reaction.

In vitro effect of medicinal plants used to treat erectile dysfunction on smooth muscle relaxation and human sperm.

Chloroform and ethanol extracts of root bark of Securidaca longepedunculata, Wrightia natalensis and Rhoicissus tridentata were investigated for their in vitro activity on the contraction of corpus cavernosal smooth muscle of white New Zealand rabbits. Some of the extracts of these plants relaxed the corpus cavernosal smooth muscle at low concentrations. The highest activity was obtained from Securidaca longepedunculata chloroform extracts at a concentration of 13.0 mg/ml, which induced 66.6% relaxation. Viagra was used as a positive control in this study. Extracts of Securidaca longepedunculata added to human spermatozoa affected certain sperm parameters negatively at 6.5 mg/ml and higher whilst there was no effect at 1.0 mg/ml.

Disordered recognition of facial identity and emotions in three Asperger type autists.

In this report we aim to explore severe deficits in facial affect recognition in three boys all of whom meet the criteria of Asperger's syndrome (AS), as well as overt prosopagnosia in one (B) and covert prosopagnosia in the remaining two (C and D). Subject B, with a familially-based talent of being highly gifted in physics and mathematics, showed no interest in people, a quasi complete lack of comprehension of emotions, and very poor emotional reactivity. The marked neuropsychological deficits were a moderate prosopagnosia and severely disordered recognition of facial emotions, gender and age. Expressive facial emotion, whole body psychomotor expression and speech prosody were quasi absent as well. In all three boys these facial processing deficits were more or less isolated, and general visuospatial functions, attention, formal language and scholastic performances were normal or even highly developed with the exception of deficient gestalt perception in B. We consider the deficient facial emotion perception as an important pathogenetic symptom for the autistic behaviour in the three boys. Prosopagnosia, the absent facial and bodily expression, and speech prosody were important but varying co-morbid disorders. The total clinical picture of non-verbal disordered communication is a complex of predominantly bilateral and/or right hemisphere cortical deficits. Moreover, in B, insensitivity to pain, smells, noises and internal bodily feelings suggested a more general emotional anaesthesia and/or a deficient means of expression. It is possible that a limbic component might be involved, thus making affective appreciation also deficient.

Expression of GnRH receptor in mouse and rat testicular germ cells.

The expression of gonadotrophin-releasing hormone receptor (GnRH-R) in germinal cells of mouse testis, whole testes, pituitary glands, and mouse ovaries was determined by means of Northern hybridization using a mouse GnRH-R [(32)P]-labelled cDNA probe. Also, the expression of GnRH-R in rat germinal cells, testis and pituitary gland was determined by Northern blot analysis using the same mouse-specific probe. Three receptor transcripts were detected in all cases. In mouse pituitary, ovary and testis, we found two GnRH-R transcripts in the proximity of 4.6-4.7 and 3.4 kb, as well as a 1.6 kb transcript in the pituitary and a 2.0-2.1 kb transcript in both the ovary and testis. Mouse germ cells also exhibited three GnRH-R transcripts of 4.7, 3.5 and 2.2 kb. Two distinct GnRH-R transcripts were also detected in the rat pituitary (4.6 and 2.1 kb), testis (4. 7 and 3.5 kb) and germ cells (4.5 and 3.5 kb). In addition, a third transcript was detected in rat pituitary (1.9 kb) and in rat testis and germinal cells (2.1 kb). The present study demonstrates that GnRH-R mRNA is expressed in rat and mouse testicular germ cells. We suggest that GnRH-R present in these cells may interact with GnRH or GnRH-like peptides produced in the testis and may be part of a paracrine system. The presence of multiple GnRH-R encoding transcripts is also of interest and warrants further studies to evaluate their regulation and function.

The influence of sera, follicular fluids and seminal plasma on human sperm-zona pellucida binding.

The purpose of this study was to assess the influence of male, female and fetal cord sera, follicular fluid, and seminal plasma on human sperm-zona pellucida binding, using the hemizona assay. Steroids, gonadotrophins, growth hormone and prolactin concentrations in follicular fluid and sera were also analysed. The influence of follicular fluid (10 or 50%, v/v) and sera (10%) on sperm-zona pellucida binding was investigated by supplementing the sperm processing medium as well as the sperm-hemizona incubation medium. Different seminal plasma concentrations (1 or 10%) were added to the sperm-hemizona incubation medium. Supplementation with 10% day 3 donor serum was used as a control throughout experimentation. Although supplementation with male sera and fetal cord serum exerted a stimulatory effect (36 and 90% respectively; P < 0.029) on sperm-zona pellucida binding, hemizona indices obtained with addition of male sera, fetal cord serum and sera obtained from sub-fertile in-vitro fertilization (IVF) patients on day 12 of their menstrual cycle did not differ significantly (P > 0.05). Final progesterone concentrations in sperm-zona pellucida incubation media (10% follicular fluid supplementation), which ranged from 0.788 to 3.85 microg/ml, enhanced sperm binding to the zonae by >100% (P < 0.02). The utilization of follicular fluid (10%) as a natural physiological stimulus to enhance sperm-zona pellucida binding in an IVF setting is recommended. The presence of seminal plasma in the spermzona pellucida incubation media showed no beneficial effect on the binding ability of sperm, and can be viewed as an unfavourable substance in the proximity of the oocyte.

The detection of blood contamination in human follicular fluid.

PURPOSE: The aim of this study was to compare the reliability of the methods conventionally used to identify low levels of blood contamination in human follicular fluid (hFF) as applicable in the clinical environment. METHODS: Follicular fluid (n = 339) and plasma samples (n = 20) were collected from patients (n = 138) attending the Centre for Fertility Studies, HF Verwoerd Hospital, University of Pretoria, South Africa. hFF blood contamination was assessed by means of (a) visual inspection, (b) hematocrit (Hct), (c) spectrophotometric analysis, (d) spectrophotometric hemoglobin kit, and (e) Combur-9-test urine sticks. RESULTS: (1) Neither hematocrit nor spectrophotometry provided reliable detection at low levels of blood contamination. (2) Visual inspection presented with a better discriminatory ability than either Hct or spectrophotometry. (3) Combur-9-test sticks identified up to 50% of blood-contaminated fluids. (4) Spectrophotometrically determined hemoglobin levels presented with weak discriminatory abilities for detecting blood-contaminated fluids. CONCLUSIONS: Visual inspection as performed in this study provides a fast and relatively reliable method for the determination of blood-contaminated hFFs. In a laboratory environment, however, it would be recommended that a combination of visual inspection, Hct, and spectrophotometric evaluation be employed for the selection of blood-free fluids.