Age-related expression of prominent regulatory elements in mouse brain: catastrophic decline of FOXO3a.

Aging is associated with changes in regulation, particularly among diverse regulators in the brain. We assayed prominent regulatory elements in mouse brain to explore their relationship to one another, stress, and aging. Notably, unphosphorylated (activated) forkhead transcription factor 3a (uFOXO3a) expressed exponential decline congruent with increasing age-related mortality. Decline in uFOXO3a would impact homeostasis, aging rate, stress resistance, and mortality. We also examined other regulators associated with aging and FOXO3a: protein kinase B (PKB), the mechanistic target of rapamycin (mTOR), 70 kDa ribosomal S6 kinase (P70S6K), and 5' AMP-activated protein kinase (AMPK). It would require powerful regulatory distortion, conflicting tradeoffs and/or significant damage to inflict exponential decline of a transcription factor as crucial as FOXO3a. No other regulator examined expressed an exponential pattern congruent with aging. PKB was strongly associated with decreases in uFOXO3a, but the aging pattern of PKB did not support a causal linkage. Although mTOR expressed a trend for age-related increase, this was not significant. We considered that the mTOR downstream element, P70S6K, might suppress FOXO3a, but remarkably, it expressed a strong positive association. The age-related pattern of AMPK was also incompatible. Literature suggested the immunological regulator NFĸB (nuclear factor kappa-light-chain-enhancer of activated B cells) increases with age and suppresses FOXO3a. This would inhibit apoptosis, autophagy, mitophagy, proteostasis, detoxification, antioxidants, chaperones, and DNA repair, thus exacerbating aging. We conclude that a key aspect of aging involves distortion of key regulators in the brain.

A multi-ingredient dietary supplement abolishes large-scale brain cell loss, improves sensory function, and prevents neuronal atrophy in aging mice.

Transgenic growth hormone mice (TGM) are a recognized model of accelerated aging with characteristics including chronic oxidative stress, reduced longevity, mitochondrial dysfunction, insulin resistance, muscle wasting, and elevated inflammatory processes. Growth hormone/IGF-1 activate the Target of Rapamycin known to promote aging. TGM particularly express severe cognitive decline. We previously reported that a multi-ingredient dietary supplement (MDS) designed to offset five mechanisms associated with aging extended longevity, ameliorated cognitive deterioration and significantly reduced age-related physical deterioration in both normal mice and TGM. Here we report that TGM lose more than 50% of cells in midbrain regions, including the cerebellum and olfactory bulb. This is comparable to severe Alzheimer's disease and likely explains their striking age-related cognitive impairment. We also demonstrate that the MDS completely abrogates this severe brain cell loss, reverses cognitive decline and augments sensory and motor function in aged mice. Additionally, histological examination of retinal structure revealed markers consistent with higher numbers of photoreceptor cells in aging and supplemented mice. We know of no other treatment with such efficacy, highlighting the potential for prevention or amelioration of human neuropathologies that are similarly associated with oxidative stress, inflammation and cellular dysfunction. Environ. Mol. Mutagen. 57:382-404, 2016. © 2016 Wiley Periodicals, Inc.

The effect of phosphatidylinositol-3 kinase inhibition on matrix metalloproteinase-9 and reactive oxygen species release from chronic obstructive pulmonary disease neutrophils.

BACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) is characterised by increased neutrophilic inflammation. A potential novel anti-inflammatory target in COPD is phosphatidylinositol-3 kinase (PI3 kinase), which targets neutrophil function. This study evaluated the effects of selective PI3Kδ inhibition on COPD blood and sputum neutrophils both in the stable state and during exacerbations. METHODS: Blood and sputum neutrophils from stable and exacerbating COPD patients were cultured with the corticosteroid dexamethasone, a pan PI3 kinase inhibitor (ZSTK474), a δ selective PI3 kinase inhibitor (GSK045) and a p38 mitogen activated protein (MAP) kinase inhibitor (BIRB 796); matrix metalloproteinase (MMP)-9 and reactive oxygen species (ROS) release were analysed. RESULTS: PI3Kδ inhibition significantly reduced MMP-9, intracellular ROS and extracellular ROS release from blood neutrophils (45.6%, 30.1% and 47.4% respectively; p<0.05) and intracellular ROS release from sputum neutrophils (16.6%; p<0.05) in stable patients. PI3Kδ selective inhibition significantly reduced stimulated MMP-9 (36.4%; p<0.05) and unstimulated and stimulated ROS release (12.6 and 26.7%; p<0.05) from blood neutrophils from exacerbating patients. The effects of the p38 MAP kinase inhibitor and dexamethasone in these experiments were generally lower than PI3Kδ inhibition. CONCLUSION: PI3Kδ selective inhibition is a potential strategy for targeting glucocorticoid insensitive MMP-9 and ROS secretion from COPD neutrophils, both in the stable state and during exacerbations.

Synergistic effects of diet and exercise on hippocampal function in chronically stressed mice.

Severe chronic stress can have a profoundly negative impact on the brain, affecting plasticity, neurogenesis, memory and mood. On the other hand, there are factors that upregulate neurogenesis, which include dietary antioxidants and physical activity. These factors are associated with biochemical processes that are also altered in age-related cognitive decline and dementia, such as neurotrophin expression, oxidative stress and inflammation. We exposed mice to an unpredictable series of stressors or left them undisturbed (controls). Subsets of stressed and control mice were concurrently given (1) no additional treatment, (2) a complex dietary supplement (CDS) designed to ameliorate inflammation, oxidative stress, mitochondrial dysfunction, insulin resistance and membrane integrity, (3) a running wheel in each of their home cages that permitted them to exercise, or (4) both the CDS and the running wheel for exercise. Four weeks of unpredictable stress reduced the animals' preference for saccharin, increased their adrenal weights and abolished the exercise-induced upregulation of neurogenesis that was observed in non-stressed animals. Unexpectedly, stress did not reduce hippocampal size, brain-derived neurotrophic factor (BDNF), or neurogenesis. The combination of dietary supplementation and exercise had multiple beneficial effects, as reflected in the number of doublecortin (DCX)-positive immature neurons in the dentate gyrus (DG), the sectional area of the DG and hippocampal CA1, as well as increased hippocampal BDNF messenger ribonucleic acid (mRNA) and serum vascular endothelial growth factor (VEGF) levels. In contrast, these benefits were not observed in chronically stressed animals exposed to either dietary supplementation or exercise alone. These findings could have important clinical implications for those suffering from chronic stress-related disorders such as major depression.

The need for training frameworks and scientific evidence in developing scarless surgery: a national survey of surgeons' opinions on single port laparoscopic surgery.

INTRODUCTION: Single port/incision laparoscopic surgery (SPILS) is a recent innovation in minimally invasive surgery whereby operations are performed through a single point of entry. Despite the relative paucity of clinical data, the procedure is increasingly being used to minimise scarring and pain associated with the multiple entry points of traditional laparoscopic surgery. This study aimed to analyse the awareness, experience and opinions of British surgeons regarding SPILS. METHODS: Electronic, 13-item, self-administered, anonymous questionnaire survey distributed via national/regional surgical mailing lists and websites. Results were collated and analysed with SPSS v17.0 for Windows (SPSS, Inc, Chicago, IL). RESULTS: 342 fully completed responses received, including 72 (21%) Consultants and 189 (55%) higher surgical trainees. Overall 330 (96.5%) were aware of SPILS; there was no significant difference in awareness between grades. Only 37% had assisted or performed SPILS procedures. More consultants performed these than trainees (56.3 vs 32.0%, p < 0.05). Operative experience was limited, with only 6% of those undertaking SPILS performing ≥25 procedures, and 60% performing ≤5. 61.4% believed SPILS takes longer to perform, and 32.8% believed it has higher complication rate. Factors cited as limiting uptake included: lack of evidence (70%), insufficient training opportunities (78%), incorrect instrumentation (70%), increased cost (62%), and hospital policy (44.5%). Patient preference was considered to have negatively affected SPILS uptake by only 9% of respondents. A greater proportion of trainees (94.6% vs 78.9%) felt there were insufficient SPILS training opportunities (p = 0.001). CONCLUSIONS: Although awareness of SPILS is high, operative experience is limited and negative perceptions regarding operating time and complications remain. The findings suggest future uptake relies strongly on the availability of evidence, training, instrumentation and reduced costs. Scientific studies are still awaited to assess effectiveness and provide clinical and economic evaluation.

Molecular epidemiology of Giardia duodenalis in an endangered carnivore--the African painted dog.

The African painted dog (Lycaon pictus) is an endangered carnivore of sub-Saharan Africa. To assist in conservation efforts a parasitological survey was conducted on wild and captive populations. Faecal samples were collected and examined for the presence of parasites using traditional microscopy techniques. The protozoan Giardia duodenalis was identified at a prevalence of approximately 26% in the wild populations and 62% in the captive population. Molecular characterisation of these isolates using three loci, 18S rRNA, ß-giardin and the glutamate dehydrogenase gene revealed the zoonotic assemblages A and B existed in high proportions in both populations. The dog assemblages C and D were rarely observed. The identification of the zoonotic genotype suggests this species has the potential to act as a reservoir for human infections. Zoonotic transmission may be possible in captive populations due to the close interaction with humans however, in wild populations anthropozoonotic transmission seems more likely. This study is the first to observe G. duodenalis in the African painted dog and to identify a possible emerging disease in this wild carnivore.

Femoral cement pressurisation for hip arthroplasty in previously fixated hips: an experimental study.

INTRODUCTION: It is common to use a cemented total hip replacement following failed hip screw fixation of a fractured femoral neck; this solution, however, is complicated by the presence of the holes that are left in the femur when the screws are removed. These holes can allow cement to leak out while being pressurised. The aim of this study was to look at the cement femoral pressures proximally and distally in a sawbone model with pre-drilled holes to assess if the commonest surgical technique of occluding the holes with fingers could maintain the cement pressure high enough. MATERIALS AND METHODS: We used eight synthetic proximal femurs, four with dynamic hip screw holes drilled in them on the lateral surface ("drilled femurs") and four with no holes ("undrilled femurs"). We used pressure sensors positioned in holes drilled in the proximal and distal parts of the medial surface to measure the pressure in the cement as it was being delivered and pressurised into the femur canal. The tests were conducted while the femur was clamped at its distal end and, in the case of the drilled femurs, while the screw holes were occluded manually. RESULTS: We found that on the proximal side, the peak cement pressure in undrilled femurs was significantly greater than in drilled femurs (p=0.006). On the distal side, the difference in peak cement pressure between the two study groups was not significant (p=0.22). At both the proximal and distal positions, the time over which the cement pressure exceeded both 5 and 100 kPa was significantly longer in undrilled femurs than in drilled femurs (p<0.05). CONCLUSION: Our results show that it is difficult to fully occlude the drill holes completely with finger tips, especially when using pressurised cement. There are significant differences in the peak cement pressures between drilled and undrilled femurs with possible consequences for patients undergoing total hip replacement.

Elevated DNA damage in a mouse model of oxidative stress: impacts of ionizing radiation and a protective dietary supplement.

Transgenic growth hormone (Tg) mice express elevated free radical processes and a progeroid syndrome of accelerated ageing. We examined bone marrow cells of Tg mice and their normal (Nr) siblings for three markers of DNA damage and assessed the impact of free radical stress using ionizing radiation. We also evaluated the radiation protection afforded by a dietary supplement that we previously demonstrated to extend longevity and reduce cognitive ageing of Nr and Tg mice. Spectral karyotyping revealed few spontaneous chromosomal aberrations in Nr or Tg. Tg mice, however, had significantly greater constitutive levels of both gammaH2AX and 8-hydroxy-deoxyguanosine (8-OHdG) compared to Nr. When exposed to a 2-Gy whole-body dose of ionizing radiation, both Nr and Tg mice showed significant increases in DNA damage. Compared to Nr mice, irradiated Tg mice had dramatically higher levels of gammaH2AX foci and double the levels of chromosomal aberrations. In unirradiated mice, the dietary supplement significantly reduced constitutive gammaH2AX and 8-OHdG in both Nr and Tg mice (normalizing both gammaH2AX and 8-OHdG in Tg), with little difference in gammaH2AX and 8-OHdG over constitutive levels. Induced chromosomal aberrations were also reduced, and in Nr mice, virtually absent. Remarkably, supplemented mice expressed 6-fold lower levels of radiation-induced chromosomal aberrations compared to unsupplemented Nr or Tg mice. Based on our data, the dietary supplement appeared to scavenge free radicals before they could cause damage. This study validates Tg mice as an exemplary model of oxidative stress and radiation hypersensitivity and documents unprecedented radioprotection by a dietary supplement comprised of ingredients available to the general public.

Studies of male reproduction in captive African wild dogs (Lycaon pictus).

Implementation of assisted breeding in the captive African wild dog is restricted by a current lack of knowledge on their reproductive physiology and the apparent difficulty of effectively manipulating the complex social dynamic of the pack in order to conduct reproductive procedures. In this study, we describe protocols for the safe and repeated capture and restraint of the African wild dog (n=7) as well as techniques for assessment of male reproductive function, semen collection and preservation. In a serendipitous finding, captive African wild dogs appeared to display significant seasonal change in male reproduction. Testicular volume and tone, spermatorrhea and the ability to collect semen by electroejaculation all increased significantly during late summer and then subsequently declined by early spring. While there were no detectable seasonal changes in testosterone concentration in the population as whole, the alpha-dominant male in both years of the study, had a highly elevated testosterone concentration compared to subordinate males. Semen collection by electroejaculation during the late summer was most effective in peri-pubertal males (15 months) when all seven electroejaculates were of adequate quality for cryopreservation. In the second breeding season (27 months), there were numerous changes in the pack hierarchy and electroejaculation was not as successful (3/7). The characteristics of electroejaculated semen collected in the breeding season are described for seven animals including the first descriptions and incidence of sperm abnormalities in the species. Semen (n=7) was frozen using a Tris-citrate fructose buffer and final egg yolk and glycerol concentration of 20% and 4%, respectively. Sperm were loaded into 0.25 mL straws, frozen in liquid nitrogen vapor and then thawed at 37 degrees C. Initial post-thaw survival of spermatozoa was encouraging (% motile: 31.8+/-5.8%; rate: 2.8+/-0.3; % intact plasma membranes: 33.4+/-5.3% and the % of damaged acrosomes: 4.4+/-1.5%) but following 2 h incubation at 37 degrees C, post-thaw survival declined markedly.

[ECG compression method by using multiple polynomial modelling: comparison with wavelet-changing technique].

The proposed ECG compression combined two approaches, ECG beat alignment and polynomial modeling. QRS complexes are firstly detected then aligned in order to reduce high frequency changes from beat to beat. These changes are modeled by means a polynomial projection. ECG from MIT-BIH database are used to evaluate the performances of the proposed technique. A comparison with the wavelet approach is performed by means the compression ratio (CR) versus the RRD curve.

A dietary supplement abolishes age-related cognitive decline in transgenic mice expressing elevated free radical processes.

We previously found that transgenic mice overexpressing growth hormone (TGM) have elevated and progressively increasing free radical processes in brain that strongly correlates with reduced survivorship. Young mature TGM, however, displayed vastly enhanced learning of an eight-choice cued maze and qualitatively different learning curves than normal controls. Here we document the age-related patterns in learning ability of TGM and normal mice. Learning appeared inferior in both genotypes of very young mice but TGM were confirmed to be superior to normal mice upon maturity. Older TGM, however, showed rapid age-related loss of their exceptional learning, whereas normal mice at 1 year of age showed little change. The cognitive decline of TGM was abolished by a complex "anti-aging" dietary supplement formulated to promote membrane and mitochondrial integrity, increase insulin sensitivity, reduce reactive oxygen and nitrogen species, and ameliorate inflammation. Results are discussed in the context of reactive oxygen and nitrogen species, long-term potentiation, learning, aging and neuropathology, based on known impacts of the growth hormone axis on the brain, and characteristics of TGM.

Technique for fabricating a lightweight, urethane-lined silicone orbital prosthesis.

A technique is presented for packing an acrylic resin core and controlling the thickness of an orbital prosthesis with the use of an irreversible hydrocolloid matrix. This technique provides a durable core without damaging the orbital prosthesis mold and permits a controllable thickness and therefore a lighter prosthesis.

Extraoral maxillofacial prosthetic rehabilitation at the M. D. Anderson Cancer Center: a survey of patient attitudes and opinions.

STATEMENT OF PROBLEM: Measures of satisfaction after extraoral maxillofacial prosthetic rehabilitation have been sparsely reported. PURPOSE: This article reviews the care-related opinions of patients who were provided extraoral prostheses at the M. D. Anderson Cancer Center over the course of 10 years. MATERIAL AND METHODS: A questionnaire was delivered to 263 extraoral prosthetic patients to elucidate their degree of satisfaction with several parameters, including prosthetic use, care, quality, durability, longevity, and cost. In addition, issues relating to self-image, socialization frequency, and income-earning ability before and after rehabilitation were surveyed. RESULTS: The views of 76 respondents demonstrated a general satisfaction with their prostheses. A majority believed that their prostheses fit comfortably, and most were satisfied with cosmesis. In addition, a preponderance of respondents reported no substantial alteration in social activity after rehabilitation. Although the number of respondents relying on their own employment fell by more than 50% after rehabilitation, nearly all reported a relatively static income before and after rehabilitation. CONCLUSION: The survey revealed a high degree of patient satisfaction with extraoral maxillofacial prostheses. Nevertheless, areas of potential improvement were not lost on the survey's population. The patients desired prostheses that last longer and have improved color stability. An interest in improved retentive mechanisms was mentioned by several patients, and only about half of the respondents perceived prosthetic cost to be completely reasonable.

Lead time reduction: the catalyst for world-class results.

With the objective of reducing lead times as much as possible, many process improvements can be made. Without the objective of lead time reduction, there can be many confusing objectives without a common thread. This usually results in poor achievement scores on many objectives. To accomplish one major objective, lead time reduction, all other areas will and must be addressed but by a united work force working towards one common goal.

Experience with Barton button and peristomal breathing valve attachments for hands-free tracheoesophageal speech.

BACKGROUND: Tracheostoma breathing valves permit hands-free tracheoesophageal (TE) speech production; however, few laryngectomees routinely use them because of problems with attachment. METHODS: We retrospectively reviewed the charts of 45 TE speakers to determine the success rate and factors associated with successful breathing valve use based on attachment. All patients attempted to use a tracheostoma breathing valve with either a standard or customized peristomal housing, or a standard or customized Barton button. Device selection was based on inspection of the patient's neck and peristomal contour. Six to eight consecutive hours of attachment defined success. RESULTS: Overall, 9% of subjects succeeded with any peristomal attachment as compared to 68% with either a standard (57%) or customized (85%) Barton button. Smooth stomal contour, a contiguous stomal lip, and correct button length were important for successful Barton button use. CONCLUSIONS: Standard or customized Barton buttons offer excellent alternatives to peristomal housing attachments for hands-free TE speech in select patients.

The use of 0.25% isoflurane premixed in 50% nitrous oxide and oxygen for pain relief in labour.

The addition of 0.25% isoflurane to 50% nitrous oxide in oxygen provides more effective pain relief in labour than 50% nitrous oxide alone. This study was carried out to determine whether self-administration by demand valve of 0.25% isoflurane in 50% nitrous oxide in oxygen premixed in cylinders at 13.7 MPa (IN2O) was practical and safe during labour. Two hundred and twenty-one mothers used IN2O in labour after 50% nitrous oxide had become inadequate for pain relief. Data on IN2O use was recorded during labour and details of the course of labour and opioid usage were taken from the clinical notes. The duration of IN2O use was 0.1-12.35 h (median 2.3). Thirty-two mothers (14.5%) required an epidural and intolerance to IN2O was seen in a maximum of 17 cases (7.7%). One hundred and twenty-six cases were primiparous and 93 parous with 151 deliveries being spontaneous and 70 interventional, of which 12 were by Caesarean section. Maternal blood loss was 20-1500 ml (median 200 ml). Apgar scores at 1 and 5 min were unaffected by IN2O use although a positive correlation was found between the use of opioids and the number of neonates with a 1-min score below 8 and the number requiring resuscitation. Six neonates had an Apgar score below 8 at 5 min, but their condition was adequately explained by factors other than the sedative technique used. Self-administered IN2O was found to be a safe and practical technique for sedation in labour when 50% nitrous oxide alone had become inadequate.

Effects of group CBT on the survival time of patients with metastatic breast cancer.

We have recently reported the psychological outcome of a group Cognitive Behaviour Therapy (CBT) intervention with patients who had metastatic breast cancer. The data of 92 patients who were retained at the first follow-up assessment revealed short-term improvements on measures of mood and self-esteem amongst therapy participants. These changes were not sustained at the 3- and 6-month follow-up assessments. This report describes a survival analysis of 121 patients who entered the study, at 5 years after its commencement. The analysis, based on the Cox Proportional Hazards Regression model, revealed no survival advantage associated with the intervention. Only medical prognostic factors such as Eastern Cooperative Oncology Group (ECOG) performance status, visceral metastases and chemotherapy treatment significantly predicted patients' survival time. The findings of this study are compared with those of two earlier studies which also examined the survival effects of a group psychological intervention with metastatic breast cancer patients.

Guided tissue fabrication from periosteum using preformed biodegradable polymer scaffolds.

A successful tissue engineering method for bone replacement would imitate natural bone graft by providing the essential elements for new bone formation using synthetic scaffolds, osteogenic cell populations, and bone induction factors. This is a study of the suitability of various formulations of poly(DL-lactic-co-glycolic acid) (PLGA) foams to provide a tissue conducting scaffold in an ovine model for bone flap fabrication. Three formulations were used of different copolymer ratio and molecular weight. Porous wafers of PLGA were stacked into rectangular chambers (volume 4 cm3) enclosed on five sides. Some chambers also contained autologous morcellized bone graft (MBG). The chambers were inserted with the open face adjacent to the cambium layer of the periosteum in rib beds of seven sheep and harvested after 8 weeks in vivo. Gross and histologic examination of the resulting tissue specimens demonstrated molded units of vascularized tissue generally conforming to the shape of the chambers and firmly attached to the periosteum. Polymer degradation appeared to occur by varying degrees based on polymer formulation. New bone formation was observed only in areas containing MBG. There was no evidence of significant inflammatory reaction or local tissue damage at 8 weeks. We conclude that a PLGA foam scaffold is (1) an efficient conductor of new tissue growth but not osteoinductive, (2) contributes to the shape of molded tissue, and (3) biocompatible when used in this model. Further studies are warranted to develop practical methods to deliver bone induction factors to the system to promote osseous tissue generation throughout the synthetic scaffold.