Non-telecentric two-photon microscopy for 3D random access mesoscale imaging.

Diffraction-limited two-photon microscopy permits minimally invasive optical monitoring of neuronal activity. However, most conventional two-photon microscopes impose significant constraints on the size of the imaging field-of-view and the specific shape of the effective excitation volume, thus limiting the scope of biological questions that can be addressed and the information obtainable. Here, employing a non-telecentric optical design, we present a low-cost, easily implemented and flexible solution to address these limitations, offering a several-fold expanded three-dimensional field of view. Moreover, rapid laser-focus control via an electrically tunable lens allows near-simultaneous imaging of remote regions separated in three dimensions and permits the bending of imaging planes to follow natural curvatures in biological structures. Crucially, our core design is readily implemented (and reversed) within a matter of hours, making it highly suitable as a base platform for further development. We demonstrate the application of our system for imaging neuronal activity in a variety of examples in zebrafish, mice and fruit flies.

LED Zappelin': An open source LED controller for arbitrary spectrum visual stimulation and optogenetics during 2-photon imaging.

Two-photon (2P) microscopy is a cornerstone technique in neuroscience research. However, combining 2P imaging with spectrally arbitrary light stimulation can be challenging due to crosstalk between stimulation light and fluorescence detection. To overcome this limitation, we present a simple and low-cost electronic solution based on an ESP32 microcontroller and a TLC5947 LED driver to rapidly time-interleave stimulation and detection epochs during scans. Implemented for less than $100, our design can independently drive up to 24 arbitrary spectrum LEDs to meet user requirements. We demonstrate the utility of our stimulator for colour vision experiments on the in vivo tetrachromatic zebrafish retina and for optogenetic circuit mapping in Drosophila.

The effects of body position on the distribution of obstructive, mixed and central sleep apnoea.

BACKGROUND: Obstructive sleep apnoea is commonly aggravated by the supine body position. The impact of body position on the severity of mixed and central sleep apnoeas is understudied. OBJECTIVES: To evaluate the impact of body position on obstructive, mixed and central apnoea indices in subjects presenting with this triform of sleep apnoea during a single polysomnogram. METHODS: We retrospectively analysed 26 polysomnograms where obstructive, mixed and central apnoeas each occurred at a rate >5/hr. Comparisons between lateral and supine body positions were made for obstructive apnoea index (OAI), mixed apnoea index (MAI), central apnoea index (CAI), apnoea-hypopnoea index (AHI) and obstructive apnoea-hypopnoea index (OAHI). RESULTS: Mean (SD) apnoea indices were significantly lower in lateral v. supine positions, respectively: MAI 15.06 (18.34) v. 32.09 (17.05); p<0.001, CAI 11.82 (11.77) v. 23.82 (14.18); p<0.001, AHI 79.46 (31.17) v. 99.47 (26.33); p<0.001, OAHI 67.87 (28.25) v. 76.00 (23.21); p=0.039. Unexpectedly, the converse was seen for OAI when comparing the lateral v. supine position: 53.10 (30.64) v. 43.58 (25.83); p=0.009, respectively. CONCLUSION: It may be beneficial for subjects with a combination of obstructive, mixed, and central apnoeas to avoid the supine body position. In this triform phenotype, mixed apnoeas are neither purely obstructive nor purely centrally mediated. Furthermore, obstructive, mixed, and central apnoeas may be different representations of a single respiratory abnormality.

Bladder stones in patients with spinal cord injury: a long-term study.

STUDY DESIGN: Retrospective follow-up study. OBJECTIVES: To assess the occurrence of bladder stones in patients with spinal cord injury (SCI). SETTING: Single SCI rehabilitation center in Switzerland. METHODS: We searched our database for SCI patients who had undergone surgery due to bladder stones between 2004 and 2012. In all patients retrieved, personal characteristics, bladder management, bladder stone occurrence and time to stone formation/recurrence were recorded. RESULTS: We identified 93 (3.3%) of 2825 patients with bladder stones, 24 women and 69 men, with a mean age 50 years (17-83) years. We observed bladder stones in patients with suprapubic catheter (SPC) in 11% (50/453), transurethral catheter (TC) in 6.6% (5/75), with intermittent catheterization (IC) in 2% (27/1315) and with reflex micturition (RM) in 1.1% (11/982), respectively. The mean time period to stone development was 95 months. The TC group had the shortest time interval (31 months), followed by the SPC group (59 months), individuals performing IC (116 months) and RM (211 months), respectively. Bladder stone recurrence rate was 23%. Recurrences were most frequent in the TC group (40%), followed by SPC (28%) and IC (22%), whereas no recurrences occurred in the RM group. Time to recurrence was shortest in the SPC group (14 months), followed by the IC (26 months) and the TC group (31 months), respectively. CONCLUSION: In SCI patients, bladder management has an important role in the development of bladder stones. Indwelling catheters (TC/SPC) are associated with the highest risk to develop bladder stones and therefore should be avoided if possible. If unavoidable, SPC are superior to TC.

[Squamous cell carcinoma of the bladder in a patient with HIV and paraplegia]. / Plattenepithelkarzinom der Harnblase bei einem Patienten mit HIV und Paraplegie.

In patients with spinal cord injury (SCI) the rate of squamous cell carcinomas (SCC) among bladder tumors is increased compared to the general population. An increased life expectancy is achieved by modern HIV treatment so that more AIDS-unrelated malignomas, e.g. bladder tumors, occur in these patients. Therefore, the risk for SCC in this group of patients is increased in patients with SCI and HIV but the combination of these two diseases is rare. We report the first case of SCC in a patient with SCI and HIV. Initial symptoms of bladder tumors in patients with SCI are often unspecific; therefore, in cases with new onset hematuria, recurrent urinary tract infections and changes in bladder function, cystoscopy and computed tomography (CT) scanning should be considered.

Residual urine volumes after intermittent catheterization in men with spinal cord injury.

STUDY DESIGN: Prospective cross-sectional study. OBJECTIVES: To investigate residual urine volumes after intermittent catheterization (IC) in men with spinal cord injury (SCI) and the effect of residual urine on the rate of symptomatic urinary tract infections (UTIs). SETTING: Single SCI rehabilitation center in Switzerland. METHODS: Sixty men experienced in IC (≥ 6 months) emptied their bladder twice by IC. Immediately after catheterization, residual urine was determined by ultrasonography. Personal characteristics and bladder diary details (annual UTI rate, catheter type) were also collected. RESULTS: The median residual urine volume was 7.0 ml (lower quartile (LQ): 0.0, upper quartile (UQ): 20.3 ml). No residual urine was observed after 42% (n=50) of all catheterizations (n=120). Unsatisfactory residual volumes (that is, >50 ml) were observed after 9% (n=11) of all catheterizations. There was no significant (P=0.95) difference between the median residual urine volume of men with recurrent (>2 UTIs per year) UTIs (2.5 ml, LQ: 0.0, UQ: 29.3 ml) and the volume of those with sporadic (≤ 2 UTIs per year) UTIs (6.0 ml, LQ: 0.0, UQ: 20.0 ml). CONCLUSIONS: Bladder evacuation by IC is an efficient method, resulting in zero or small residual urine volumes. The small residual urine volumes generally observed after IC do not predispose for UTIs.

[Urethral condyloma in a patient with neurogenic bladder dysfunction: a therapeutic challenge]. / Urethrale Kondylome bei einem Patienten mit neurogener Blasenfunktionsstörung : Eine therapeutische Herausforderung.

Urethral condyloma is a therapeutic challenge. This article reports the case of a patient with spinal cord injury with the incidental finding of a massive spread of urethral condyloma. After removal of the condyloma with biopsy forceps neither recurrence of the condyloma nor a urethral stricture occurred. In patients where intermittent catheterization is performed, condyloma may be dispersed into the bladder, therefore, prompt endoscopic removal is crucial. As condyloma frequently recurs even after complete removal regular controls are mandatory.

[A cylinder aneurysm of a penile prosthesis]. / Das Penisprothesenaneurysma.

Herniation of a penile prosthesis (cylinder aneurysm) is an extremely rare complication of penile prosthesis surgery. We report the first case of such an aneurysm in a patient with spinal cord injury. The treatment of choice is surgical revision with replacement of the faulty device. Filling of the implanted system with contrast media facilitates preoperative diagnostic workup.

Gemcitabine, paclitaxel, and cisplatin as combined adjuvant approach in transitional cell carcinoma of the urothelium.

AIM: The objective of this study was to evaluate the adjuvant chemotherapy with gemcitabine, paclitaxel, and cisplatin for urothelial carcinoma. METHODS: Twenty-seven patients with invasive transitional cell carcinoma of the urothelium were treated between 2001 and 2007. All patients received chemotherapy with intravenous gemcitabine at a dose of 1000 mg/mC on Days 1 and 8, intravenous paclitaxel at a dose of 80 mg/mC on Days 1 and 8, and intravenous cisplatin at a dose of 50 mg/mC on Day 2. Treatment courses were repeated every 21 days. RESULTS: Median follow-up period was 32.5 months. Six patients came to progressive disease. The median overall survival was not reached, and the actuarial 1-year and 2-year survival rates were 89% and 67% respectively. The median progression-free survival was 10.0 months. Median survival time for patients with ECOG status 0, and 1 was 52.0, and 22.0 months respectively. Grade 4 neutropenia occurred in 18.5% of patients, but there was no treatment related mortality. CONCLUSIONS: The combination of gemcitabine, paclitaxel, and cisplatin is a highly effective and tolerable regimen for patients with invasive transitional cell carcinoma of the urothelium. This treatment should be considered as a suitable option that deserves further prospective evaluation. ECOG performance status is an important predictive factors for survival.

Palliative chemotherapy with gemcitabine, paclitaxel, and cisplatin as first-line treatment following gemcitabine monotherapy for patients with transitional cell carcinoma of the urothelium.

AIM: Chemotherapeutic agents are active in transitional cell cancer of the urothelium, and combinations have shown promising results. The objective of this study was to evaluate the palliative chemotherapy with gemcitabine, paclitaxel, and cisplatin for transitional cell carcinoma. METHODS: Thirty-four patients with advanced transitional cell carcinoma of the urothelium were treated between 2000 and 2007. All patients received chemotherapy with intravenous gemcitabine at a dose of 1000 mg/m2 on days I and VIII, intravenous paclitaxel at a dose of 80 mg/m2 on days I and VIII, and intravenous cisplatin at a dose of 50 mg/m2 on day II. Treatment courses were repeated every 21 days. After completion of four to six courses in this regimen an application of intravenous gemcitabine at a dose of 1000 mg/m2 followed repeating every 28 days. RESULTS: Twelve patients (35.3%) had 1 visceral sites of metastases. Twenty two patients (64.7%) had achieved objective responses to treatment (29.4% complete responses). The median actuarial survival was 18.5 months, and the actuarial one-year and two-year survival rates were 56% and 26% respectively. After a median follow-up of 16.3 months, 18 patients remained alive. The median progression-free survival was 7 months. Median survival time for patients with ECOG status 0, 1, and 2 was 45, 12, and 10.5 months respectively. Grade 3-4 neutropenia occurred in 41.2% of patients. CONCLUSIONS: The combination of gemcitabine, paclitaxel, and cisplatin is a highly effective and tolerable regimen for patients with advanced transitional cell carcinoma of the urothelium. This treatment should be considered as a suitable option that deserves further prospective evaluation. ECOG performance status and visceral metastases are important predictive factors for survival.

Attention and working memory in resident anaesthetists after night duty: group and individual effects.

AIMS: To investigate the effects of a single period of night duty on measures of attention and working memory in a group of residents (registrars) in anaesthesiology. Emphasis was placed on individual deficits using a reference point of the equivalent effect of a blood alcohol concentration (BAC) >0.05% determined by other researchers. METHODS: There were 33 subjects aged 26-42 years. Night duty was performed on a weekly basis. Baseline assessments were conducted at either 08 15 or 08 55 preceding night duty and repeated 24-25 hours later, just after the completion of duty. Questionnaires included items regarding duration of sleep and the Stanford Sleepiness Scale. A battery of four reaction time (RT) tasks of increasing difficulty, lasting approximately 35 minutes, was administered on a personal computer. These ranged from simple RT to progressively more complex RT tasks incorporating working memory. A significant change was regarded as >15% deterioration in respect of speed or accuracy. RESULTS: The mean duration of sleep preceding night duty was 7.04 hours and 1.66 hours during the period of night duty. Intergroup comparisons revealed significant prolongation in mean response speed in the first three tests. Mean accuracy was significantly reduced only in respect of the two more complex tests. A >15% deterioration in response speed occurred in up to 30% of subjects on a single task, rising to 52% (17/33) overall. Deterioration occurred in a patchy distribution in most subjects, involving no more than one or two of the four tasks. As regards accuracy, the prevalence of deterioration increased with task complexity. CONCLUSIONS: Results are in general agreement with previous group analyses. A new dimension was added by the analysis of a broad spectrum of individual response to sleep deprivation. The effects of sleep loss in residents cannot be overlooked, even in a relatively benign work schedule.

Aging effects on vernier hyperacuity: a function of oscillation rate but not target contrast.

PURPOSE: Most previous studies have shown static vernier acuity for high-contrast targets to be largely unaffected by aging; those examining adult age differences on oscillatory displacement thresholds have consistently reported marked age deficits. The goals of this study were to (1) measure the age deficit on oscillatory discrimination beyond that attributable to any change in spatial discrimination by using the same target configuration for both task types and (2) determine whether an age-related change in the contrast response of the visual system contributes to age differences on static or oscillatory discrimination. METHODS: The displacement thresholds of young and old observers for a vernier task configuration were determined at two target contrast levels (0.08 and 0.64) for static and oscillating targets (2 and 6 Hz) RESULTS: No age differences were seen on static displacement thresholds at either high or low contrast. A marked age deficit that emerged when oscillation was increased was unrelated to target contrast or observer contrast sensitivity. CONCLUSIONS: Age-related declines in oscillatory discrimination beyond those attributable to spatial discrimination do not appear to be attributable to optical factors nor to a decline in the contrast response of the senescent visual system. These findings are discussed in terms of a functional decline in the magnocellular pathway, or "neural entropy," possibly due to random cell loss.

Vitamin A status in relation to electroretinographic and electrooculographic findings in HIV infection.

OBJECTIVES: To investigate the electrooculogram (EOG) and an expanded battery of electroretinograms (ERGs) in HIV-infected subjects without retinopathy at two different levels of immunosuppression. Consideration was given to the possible effects of serum levels of vitamin A. METHODS: Two groups of neurologically normal HIV-positive subjects with CD4+ cell counts of <200 (n=17) or >400 (n=19) were contrasted with 20 HIV-negative controls. Rod ERGs, oscillatory potentials and cone ERGs were obtained using ganzfeld stimulation. A sequence of dark- and light-adapted EOG recordings was analyzed to obtain light/dark ratios. The HIV-positive subjects underwent a comprehensive ophthalmologic examination. RESULTS: Intergroup comparisons failed to reveal any significant intergroup differences. Abnormal ERG findings, those exceeding the 98% confidence limits, occurred in 11% (4/36) of the HIV-positive subjects. One of these subjects was hyporetinemic and another had a subnormal retinol level. No subjects had abnormal EOGs. CONCLUSIONS: HIV infection influenced retinal function in 11% of a group of neurologically normal subjects without clinical signs of retinopathy. The profile of electrophysiological findings suggested predominant functional deficits involving middle or inner retinal layers. Vitamin A deficiency and possible vascular changes could account for the observed ERG abnormalities.

Threonine phosphorylation of the MMAC1/PTEN PDZ binding domain both inhibits and stimulates PDZ binding.

Two-hybrid searches with the tumor suppressor MMAC1/PTEN isolated the proteins hDLG and hMAST205. Further two-hybrid analysis and microtiter plate binding assays localized the sites of interaction to PDZ domains from hDLG and hMAST205 and the PDZ binding domain at the COOH terminus of MMAC1/PTEN. A synthetic peptide derived from the MMAC1/PTEN PDZ binding domain (MMAC1/PTEN-PDZBD) was used to coprecipitate proteins from A431 human cell lysate. The recovered proteins were resolved by SDS-PAGE and immobilized on a nitrocellulose membrane. Treatment of this membrane with an anti-hDLG antibody identified a Mr 140,000 band, consistent with the size of hDLG. Treatment of this membrane with the MMAC1/PTEN-PDZBD peptide identified a single prominent band of slightly larger than Mr 200,000 (Mr 200,000 kDa). Threonine phosphorylation of the MMAC1/ PTEN-PDZBD peptide inhibited both microtiter plate binding to the hDLG and hMAST205 PDZ domains and coprecipitation of the Mr 140,000 and > 200,000 proteins, but promoted coprecipitation of proteins of approximately Mr 90,000 and Mr 120,000 from A431 cell lysate. This result suggests phosphorylation of the MMAC1/PTEN PDZ binding domain can both inhibit and promote PDZ interactions.

Once versus twice daily amikacin in neonates: prospective study on toxicity.

OBJECTIVE: To compare the potentially toxic effects in fullterm neonates of amikacin administered once daily, versus amikacin administered twice daily. METHODOLOGY: A controlled, randomized, prospective study in which one group of fullterm neonatal patients received amikacin 15 mg/kg per dose once daily (n = 20), and the other received amikacin 7.5 mg/kg per dose twice daily (n = 20). Impairment of renal glomerular function was defined as a decline of less than 50% of the expected physiological drop in serum creatinine over time. Brainstem auditory evoked potentials were also evaluated and amikacin blood levels taken. RESULTS: Fifteen patients in the once-daily group and 12 patients in the twice-daily group demonstrated at least one period of renal function impairment while in hospital. This decreased to five of 16 and four of 16 patients during follow-up. These differences were not statistically significant. Brainstem auditory evoked potentials did not find signs of ototoxicity at any time. CONCLUSION: In fullterm neonatal patients, once daily dosing of amikacin is no more toxic than the twice daily regimen.

Discrimination between viral and nonviral meningitis by visually analyzed and quantitative electroencephalography.

A prospective study was conducted to assess the ability of the visually analyzed electroencephalogram (VEEG), the quantitative EEG (QEEG) and the Glasgow Coma Scale (GCS) to discriminate between patients with viral and nonviral meningitis. The 55 subjects, aged 14-75 years, fell into one of the following categories: viral (n = 12), bacterial (n = 19), tuberculous (n = 16) or cryptococcal (n = 8) meningitis. EEG recordings and Glasgow Coma Scale (GCS) scores were obtained within 48 hours of admission to hospital. The sensitivity of the VEEG and QEEG for the prediction of patients with nonviral meningitis (true positives in this context) attained reasonably high values of 70% and 80%, respectively. In contrast, the sensitivity of the GCS was only 38%. Each of the three tests achieved high degrees of consistency in this regard with positive predictive values of 94% or better. The specificity for each of the three tests was high, 100% for the VEEG and the GCS and 82% for the QEEG indicating a high probability for the correct prediction of viral meningitis (true negatives). The consistency of this prediction was, however, poor due to negative predictive values of only 53% for the QEEG, 48% for the VEEG and 32% for the GCS. The QEEG results did not reveal any obvious advantages over the VEEG. Rather the assessment of the occurrence of particular VEEG abnormalities showed that patients with delta abnormalities had a very high probability of nonviral meningitis. At the other end of the spectrum, all normal VEEGs occurred in viral meningitis. In important respects the predictive ability of the EEG was superior to that of the GCS. While there was statistically significant agreement between the VEEG and GCS, the degree of agreement was poor. This study indicates that the EEG is a valuable and probably underestimated test in the acute phase of meningitis and provides complementary information to the GCS.

Characterization of a carboxy-terminal BRCA1 interacting protein.

There are several lines of evidence indicating that the carboxy-terminal region of the tumor suppressor protein BRCA1 is a functionally significant domain. Using the yeast two-hybrid and in vitro biochemical assays, we show that a protein, CtIP, interacts specifically with the carboxy-terminal segment of human BRCA1 from residues 1602-1863. A germ line truncation mutation, Y1853ter, that removes the last 11 amino acids from the carboxy-terminus of BRCA1, abolishes not only its transcriptional activation function, but also binding to CtIP. The function of CtIP is unknown, but its reported association with a transcriptional repressor CtBP lends further support that it may have a role in transcription. A sequence based screen of a panel of 89 tumor cell line cDNAs for mutations in the CtIP coding region identified five missense variants. In the pancreatic carcinoma cell line, BxPC3, the non-conservative lysine to glutamic acid change at codon 337 is accompanied with apparent loss of heterozygosity or non-expression of the wild type allele. Thus it is plausible that CtIP may itself be a tumor suppressor acting in the same pathway as BRCA1.

Human mitogen-activated protein kinase kinase 4 as a candidate tumor suppressor.

Mitogen-activated protein kinases function in signal transduction pathways that are involved in controlling key cellular processes in many organisms. A mammalian member of this kinase family, MKK4/JNKK1/SEK1, has been reported to link upstream MEKK1 to downstream stress-activated protein kinase/JNK1 and p38 mitogen-activated protein kinase. This mitogen-activated protein kinase pathway has been implicated in the signal transduction of cytokine- and stress-induced apoptosis in a variety of cell types. Here, we report that two human tumor cell lines, derived from pancreatic carcinoma and lung carcinoma, harbor homozygous deletions that eliminate coding portions of the MKK4 locus at 17p, located approximately 10 cM centromeric of p53. In addition, in a set of 88 human cancer cell lines prescreened for loss of heterozygosity, we detected two nonsense and three missense sequence variants of MKK4 in cancer cell lines derived from human pancreatic, breast, colon, and testis cells. In vitro biochemical assays revealed that, when stimulated by MEKK1, four of the five altered MKK4 proteins lacked the ability to phosphorylate stress-activated protein kinase. Thus, the incidence of coding mutations of MKK4 in the set of cell lines is 6 of 213 (approximately 3%). These findings suggest that MKK4 may function as a suppressor of tumorigenesis or metastasis in certain types of cells.

Short-term antihypertensive medication does not exacerbate sleep-disordered breathing in newly diagnosed hypertensive patients.

It has been speculated for some time that various antihypertensive medications may have a deleterious effect on respiration during sleep and thereby enhance the apparent association between hypertension and sleep apnea/hypopnea (SAH). However, there are few data to support this contention. The present study used a double-blind, randomized, cross-over design to contrast the effects of 6 weeks treatment with alpha-methyldopa and the combination of hydrochlorothiazide and amiloride with that of amlodipine and the combined diuretics in a group of 24 newly diagnosed patients with primary hypertension. All-night polysomnography was performed before the initiation of therapy (baseline) and at the end of the two treatment periods. Respiratory variables failed to reveal any significant differences between the treatments and baseline, or between the two different treatment regimens. The two treatment regimens achieved similar reductions in blood pressure. The prevalence of SAH was 25% before treatment, which is comparable to a prevalence of 20% in a similar group drawn from the same population but receiving various antihypertensive medications. The findings of this study are in agreement with previous reports using other classes of antihypertensive drugs that also failed to detect any tendency for increases in nocturnal respiratory disturbance indices over assessment periods of 8 weeks or shorter.