[Pelvic radiotherapy and artificial urinary sphincter in women]. / Radiothérapie pelvienne et sphincter artificiel urinaire chez la femme.

OBJECTIVES: A retrospective evaluation of artificial urinary sphincter (AUS) implantation in women with previous pelvic radiotherapy (PR). POPULATION AND METHODS: From May 1987 to December 2009, on the 215 women implanted with AUS, nine (4.2%) had previous PR. We compared two groups of women, the first one without PR (group 1; n=206) and the other group with PR (group 2; n=9). Previous preop. urodynamics were realized. Patients using more than one pad per day at the end of follow-up were considered in failure. RESULTS: Mean follow-up for these two groups was 6 years (SD: 5.6 years), with a mean age of 62.8 years. Mean delay between PR and surgery was 14 years. PR was indicated for cervix cancer in 78% (7/9), endometrial cancer and ovarian cancer in 9% (1/9) each. PR was responsible of an increased rate of AUS erosion and explantation (P<0.001). In group 2, more than half of women had AUS failure and 60% for AUS erosion, versus 22% and 26% respectively in group 1. In group 2, all the AUS eroded were explanted, one third of women, with a mean delay of 59.8 months (4-140) with AUS implantation. CONCLUSION: AUS implantation in a female population with previous PR is not necessary inconsistent, but the failure rate is high. This difficult surgery should be reserved for specialized centres.

Non-anti-MAG DADS neuropathy as a variant of CIDP: clinical, electrophysiological, laboratory features and response to treatment in 10 cases.

BACKGROUND AND PURPOSE: Some patients within the spectrum of chronic inflammatory demyelinating polyradiculoneuropathies (CIDP) have distal acquired demyelinating symmetric (DADS) neuropathy, usually associated with anti-myelin-associated-glycoprotein (MAG) IgM monoclonal gammopathy. The aim of this retrospective study was to investigate patients with DADS neuropathy without anti-MAG antibodies, and study their response to immunotherapy. METHODS: Patients were selected on the basis of (i) 'Definite CIDP' according to the EFNS/PNS Guideline criteria, (ii) The presence of disproportionately prolonged motor latencies resulting in a terminal latency index (TLI) ≤ 0.25 in at least two motor nerves and (iii) The absence of anti-MAG antibodies on ELISA. Response to immunotherapy was defined as persistent improvement by at least one point on the INCAT disability score. RESULTS: Data from 146 CIDP patients were analysed, and 10 patients were included. Six had clinically pure sensory neuropathy, and four had sensorimotor neuropathy. Ataxia was present in nine patients, generalized areflexia in seven and postural tremor in two. Five of the 10 patients had abnormal sensory potentials only in the upper limbs. An associated condition was found in nine patients: two chronic lymphocytic leukaemias, four IgG monoclonal gammopathies (one associated with non-Hodgkin's lymphoma) and two IgM monoclonal gammopathies of unknown significance. Patients were mostly improved with intravenous immunoglobulin (IVIg), corticosteroids, plasma exchanges, or a combination thereof. CONCLUSION: DADS neuropathy without anti-MAG antibodies is more likely to be considered a variant of CIDP. In addition, such patients should be systematically investigated for an associated haematological or immunological condition.

[Double spinofixation laparoscopic versus laparoscopic robot-assisted: Morbidity, anatomical and functional short-term results]. / Double promontofixation laparoscopique versus laparoscopique robot-assistée : morbidité, résultats anatomiques et fonctionnels à court terme.

OBJECTIVE: The purpose of our study was to compare the morbidity and the short-term anatomical and functional outcome of the double promontofixation according to the surgical access laparoscopic versus laparoscopic robot-assisted. METHODS: Forty-six patients were operated for anterior and posterior promontofixation with two mesh between March 2008 and February 2010, 19 were robot-assisted (PR) and 27 laparoscopic (PL). All the patients were contacted again by telephone to answer a questionnaire estimating the functional results. RESULTS: Both groups (PR vs PL) were comparable in terms of age, score ASA and of surgical histories. There was no difference in terms of hospital stay nor per- and postoperative complications. The mean operating time was significantly more important in the group PR (P=0.049). With a mean follow-up of 10,7±7,8 months (PL) versus 8,8±5,9 months (PR), the anatomical result was good without recurrence in 93,5% of the cases. The rate of recurrence was similar in both groups with three patients who had a cystocele grade 2, two in the group PR and one in the group PL (P=0.411). The urinary and sexual functional results were comparable between both groups with an improvement of the rate of dyspareunia and dysuria. However we observed more postoperative constipation in the group PR (10/19 vs 6/27, P=0.033). CONCLUSION: The robot-assisted laparoscopic promontofixation is a reproducible technique with a morbidity and anatomical and functional results comparable to the laparoscopic way.

A retrospective analysis of stereotactic body radiation therapy for canine heart base tumors: 26 cases.

INTRODUCTION: This study retrospectively evaluated outcomes and adverse radiation effects (AREs) associated with stereotactic body radiation therapy (SBRT) for canine heart base tumors (HBTs). A secondary aim was to identify any demographic or echocardiographic factors that might determine which dogs would most benefit from SBRT. ANIMALS: Twenty-six dogs that received SBRT for an imaging-based diagnosis of a HBT were evaluated. METHODS: Twenty-three dogs were treated with three fractions of 10 Gy delivered daily or every other day. The remaining 3 dogs received variable protocols of one to five fractions. Demographic, echocardiographic, and radiographic information, AREs, and treatment responses were collected. Correlations of these data with survival time were evaluated. RESULTS: The median overall survival time was 404 days (95% confidence interval: 239-554 days). The majority of dogs experienced a partial response (25%) or stable disease (60%) for a median duration of 333 days (95% confidence interval: 94-526 days). Three dogs had progressive disease within six months of SBRT. Radiographic pneumonitis was identified in 7 of 23 dogs, and clinical pneumonitis was identified in 4 dogs. No other AREs were noted. The rate of distant metastasis was 13%. On multivariate analysis, it was found that vena caval obstruction, supraventricular and ventricular arrhythmias, clinical signs, and enlarged locoregional lymph nodes at presentation were negatively associated with survival time. CONCLUSIONS: Stereotactic body radiation therapy was delivered with a low rate and degree of normal tissue complications. Asymptomatic dogs with confirmed, progressive growth of a HBT may most likely benefit from SBRT.

Correlations between numerical chromosomal aberrations in the tumor and peripheral blood in canine lymphoma.

Lymphoma is the most frequently diagnosed hematopoietic malignancy in dogs. Untreated, the survival times are approximately one month. Chemotherapy is the current standard of care and can initiate and temporarily maintain remission, with average remission times of one year. Canine lymphoma is an established model of human non-Hodgkin's lymphoma, and studying this disease in dogs can provide insight to both human and canine disease. Cytogenetic abnormalities can aid in diagnosing tumors as well as in giving a more accurate prognosis for the specific mutations present. Evaluating peripheral lymphocytes instead of tumor cells is less invasive for the affected dog and technically easier. This study was designed to investigate a correspondence between numerical aberrations detected in the tumor and the peripheral blood in dogs with lymphoma. Twenty-five dogs with lymphoma had one lymph node excised, a peripheral blood sample drawn, and a bone marrow aspirate performed. Portions of the lymph node were submitted for immunophenotyping and cytogenetic analysis. The peripheral blood sample was cultured for cytogenetic analysis and the bone marrow aspirate was used for staging purposes. A significant correspondence between the numerical aberrations in the tumor and the peripheral blood was found. The findings in this study pave the way toward an alternative method for evaluating lymphoma. When tumor analysis is not possible, the peripheral blood offers a viable option for cytogenetic assessment. Additionally, this may provide a method to evaluate the efficacy of the treatment protocol during the course of treatment.

Outcome of 9 dogs treated with stereotactic radiation therapy for primary or metastatic vertebral osteosarcoma.

Primary and metastatic vertebral osteosarcoma (OSA) in the dog carries an overall guarded prognosis. Previously reported definitive treatments in dogs with vertebral OSA have included surgery, radiotherapy, chemotherapy or a combination of those therapies. This retrospective study was completed to determine patterns of failure, duration of local control and survival time in dogs with vertebral OSA treated with stereotactic radiation therapy (SRT). Nine dogs were treated with SRT for vertebral OSA. Protocols ranged from 1 to 5 fractions with total prescription ranging from 13.5 to 36 Gy. Six dogs had primary lesions and 3 had metastatic lesions. Neurologic score improved in 4 patients, remained the same in 4 and worsened in 1. Five of the 6 dogs that presented with assessable spinal pain had reported improvement in pain. Overall median survival time was 139 days and median duration of pain control was 77 days. There was not a statistically significant survival difference between dogs presenting with primary or metastatic disease, or dogs that had improvement in neurologic score following SRT. The data suggests similar survival times to the previously reported definitive treatments in dogs with vertebral OSA and displays continued difficulty in controlling this tumour. The dose limiting structure is the late responding spinal cord, but many of the patients herein died prior to the expected time to development of late radiation side effects.

Impact of heterogeneity in the predictive value of kinetic parameters in canine osteosarcoma.

Intratumoral heterogeneity has been identified as a potential problem in the efficacy of predictive assays. Canine osteosarcoma is an extremely heterogeneous solid tumor that has been shown to be an excellent model for the human disease. Intratumoral heterogeneity of kinetic parameters and the effect of heterogeneity on predicting outcome of treatment (time until metastasis) were studied in dogs with naturally occurring osteosarcoma. Dogs were treated with amputation or tumor excision and limb-sparing followed by chemotherapy with cisplatin. Kinetic parameters evaluated included v, duration of DNA synthesis (Ts), and potential doubling time (Tpot), determined using in vivo labeling with bromodeoxyuridine and flow cytometry. In 30 tumors, multiple samples were obtained and evaluated. There was significantly more variation between tumors from different dogs than intratumoral variation of v, Ts, and Tpo. Variations in v, Ts, and Tpot within a tumor were associated with both sample location and tumor subpopulation. Time to metastasis was determined in 51 dogs with tumors sampled for kinetics. Multiple samples were available from 25 of these tumors. Cox proportional hazard analysis was performed using either the fastest or slowest Tpot from each sample. The fastest available Tpots were highly significant (P < 0.001) for prediction of outcome. The slowest available Tpots were also significant predictors, although the statistical strength was compromised (P = 0.024). Obtaining at least two samples in large tumors known to be heterogeneous is recommended to improve the predictive ability of Tpot. v is a more limited predictor but can useful when Tpot is not available. In canine osteosarcoma, an extremely heterogeneous tumor, kinetic parameters were shown to be predictors of outcome.

Comparison of DNA aneuploidy of primary and metastatic spontaneous canine osteosarcomas.

Spontaneous canine osteosarcomas were analyzed for DNA aneuploidy and percentage of S phase cells using flow cytometry. Forty-eight dogs were studied in which both a primary tumor and subsequent metastases were available. The DNA index distributions for the primary tumors and the metastases were quite similar. However, when individual primary tumors and metastases derived from them were compared, many of the cases had different ploidy values. The tumor cells were also analyzed for percentage of S phase. The diploid metastases had less than 17% S phase cells, whereas the aneuploid metastases had up to 40% S phase cells. There was a direct correlation between the DNA index and the percentage of S phase in the metastases.

Evaluation of 18F-FDG PET/CT as a diagnostic imaging and staging tool for feline oral squamous cell carcinoma.

18F-fluorodeoxyglucose positron emission tomography combined with computed tomography (18FDG-PET/CT) has been shown to be effective for staging human oral squamous cell carcinoma (SCC) but its application for cats with oral SCC is unknown. Twelve cats with biopsy-proven oral SCC were imaged with whole body 18FDG-PET/CT to determine its value as a diagnostic imaging and staging tool and fine needle aspirates were obtained of accessible regional lymph nodes. All tumors were FDG avid and conspicuous on 18FDG-PET/CT images, with an average of the maximum standardized uptake value 9.88 ± 5.33 SD (range 2.9-24.9). Soft tissue infiltrative tumors that were subtle and ill defined on CT were highly visible and more extensive on FDG-PET/CT. Tumors invading the osseous structures were more similar in extent on 18FDG-PET/CT and CT although they were more conspicuous on PET images. Three cytologically confirmed metastases were hypermetabolic on PET, while two of those metastases were equivocal on CT.

Stereotactic radiation therapy for treatment of canine intracranial meningiomas.

The objective of this study is to determine the rate of toxicity, median survival time (MST) and prognostic factors in dogs with presumed intracranial meningiomas that were treated with stereotactic radiation therapy (SRT). Patient demographics, neurological history, details of SRT plans and response to treatment (including toxicity and survival times) were examined for potential prognostic factors. Overall MST (MST) due to death for any cause was 561 days. There was a mild to moderate exacerbation of neurological symptoms 3-16 weeks following SRT treatments in 11/30 (36.7%) of dogs. This presumed adverse event was treated with corticosteroids, and improvement was seen in most of these dogs. Death within 6 months of treatment as a result of worsening neurologic signs was seen in 4/30 (13.3%) of dogs. Volume of normal brain that received full dose at a prescription of 8Gy × 3 fractions was predictive of death due to neurological problems within this 6-month period.

Assessment of predictive molecular variables in feline oral squamous cell carcinoma treated with stereotactic radiation therapy.

This study evaluated molecular characteristics that are potentially prognostic in cats with oral squamous cell carcinoma (SCC) that underwent stereotactic radiation therapy (SRT). Survival time (ST) and progression-free interval (PFI) were correlated with mitotic index, histopathological grades, Ki67 and epidermal growth factor receptor expressions, tumour microvascular density (MVD), and tumour oxygen tension (pO(2)). Median ST and PFI were 106 and 87 days, respectively (n = 20). Overall response rate was 38.5% with rapid improvement of clinical symptoms in many cases. Patients with higher MVD or more keratinized SCC had significantly shorter ST or PFI than patients with lower MVD or less keratinized SCC (P = 0.041 and 0.049, respectively). Females had significantly longer PFI and ST than males (P ≤ 0.016). Acute toxicities were minimal. However, treatment-related complications such as fractured mandible impacted quality of life. In conclusion, SRT alone should be considered as a palliative treatment. MVD and degree of keratinization may be useful prognostic markers.

Predicting clinical outcome in feline oral squamous cell carcinoma: tumour initiating cells, telomeres and telomerase.

Feline oral squamous cell carcinoma (SCC) has very poor prognosis. Here, a retrospective pilot study was conducted on 20 feline oral SCC patients who underwent stereotactic radiation therapy (SRT), to evaluate: (1) the value of putative tumour initiating cell (TIC) markers of human head and neck SCC (CD44, Bmi-1); (2) telomere length (TL) specifically in putative TICs; and (3) tumour relative telomerase activity (TA). Significant inverse correlations were found between treatment outcomes and Bmi-1 expression, supporting the predictive value of Bmi-1 as a negative prognostic indicator. While TL exhibited a wide range of variability, particularly in very short fractions, many tumours possessed high levels of TA, which correlated with high levels of Bmi-1, Ki67 and EGFR. Taken together, our results imply that Bmi-1 and telomerase may represent novel therapeutic targets in feline oral SCC, as their inhibition - in combination with SRT - would be expected to have beneficial treatment outcome.

Cardiovascular, neuroendocrine, and monoaminergic responses to psychological stressors: possible differences between remitted panic disorder patients and healthy controls.

Both clinical symptomatology and stress research suggest that panic attacks might be partially attributable to exaggerated psychophysiological responses to environmental stressors. In the present study, we aimed to explicitly test this idea by measuring the physiological responses to a mild psychological stressor in both healthy controls (n = 8) and fully remitted, medication-free panic disorder patients (n = 8). One hour before the stressor, former patients, compared to healthy controls, exhibited higher diastolic blood pressure. From a blood sample taken 30 min before the stressor, patients, compared to controls, had lower paroxetine platelet binding site densities. During the stressor, patients, compared to controls, had greater increases in plasma levels of cortisol. These preliminary findings suggest that remitted panic disorder patients might have disturbed physiological responses to mild psychological stressors. These disturbances might be related to the development of future episodes.

Hyperthermic treatment of malignant diseases: current status and a view toward the future.

New studies in hyperthermia at the basic science, engineering, and clinical level have stimulated renewed enthusiasm for re-investigating its potential as an anticancer therapy. This article reviews the salient features of these recent results and points out areas for additional investigation. Highlighting these new results is the publication of several positive phase III trials for thermoradiotherapy compared to radiotherapy alone. Important highlights are the encouraging results using magnetic resonance imaging for noninvasive thermometry. If this technology is successfully implemented with real time power control it will revolutionize the clinical application of hyperthermia.

Effects of intraoperative irradiation and intraoperative hyperthermia on canine sciatic nerve: neurologic and electrophysiologic study.

PURPOSE: Late radiation injury to peripheral nerve may be the limiting factor in the clinical application of intraoperative radiation therapy (IORT). The combination of IORT with intraoperative hyperthermia (IOHT) raises specific concerns regarding the effects on certain normal tissues such as peripheral nerve, which might be included in the treatment field. The objective of this study was to compare the effect of IORT alone to the effect of IORT combined with IOHT on peripheral nerve in normal beagle dogs. METHODS AND MATERIALS: Young adult beagle dogs were randomized into five groups of three to five dogs each to receive IORT doses of 16, 20, 24, 28, or 32 Gy to 5 cm of surgically exposed right sciatic nerve using 6 MeV electrons and six groups of four to five dogs each received IORT doses of 0, 12,16, 20, 24, or 28 Gy simultaneously with 44 degrees C of IOHT for 60 min. IOHT was performed using a water circulating hyperthermia device with a multichannel thermometry system on the surgically exposed sciatic nerve. Neurologic and electrophysiologic examinations were done before and monthly after treatment for 24 months. Electrophysiologic studies included electromyographic (EMG) examinations of motor function, as well as motor nerve conduction velocities studies. RESULTS: Two years after treatment, the effective dose for 50% complication (ED50) for limb paresis in dogs exposed to IORT only was 22 Gy. The ED50 for paresis in dogs exposed to IORT combined with IOHT was 15 Gy. The thermal enhancement ratio (TER) was 1.5. Electrophysiologic studies showed more prominent changes such as EMG abnormalities, decrease in conduction velocity and amplitude of the action potential, and complete conduction block in dogs that received the combination of IORT and IOHT. The latency to development of peripheral neuropathies was shorter for dogs exposed to the combined treatment. CONCLUSION: The probability of developing peripheral neuropathies in a large animal model was higher for IORT combined with IOHT, than for IORT alone. The dose required to produce the same level of late radiation injury to the sciatic nerve was reduced by a factor of 1.5 (TER) if IORT was combined with 44 degrees C of IOHT for 60 min.

Intraoperative radiotherapy with localized radioprotection: diminished duodenal toxicity with intraluminal WR2721.

The radiosensitive duodenum must be treated during IORT of human pancreatic head tumors, leading to an approximately 25% incidence of late bleeding. This study aimed to decrease the toxicity by administering WR2721 directly into the duodenal lumen. Duodenal toxicity in the canine was evaluated after intraoperative radiotherapy (IORT) with and without the intraluminal radioprotector WR2721. Eight adult dogs were divided into two groups. All underwent IORT using a 5.7 cm cone that covered the duodenum and pancreas. 30.0 Gy IORT was given with 6 MeV electrons. Cholecystojejunostomy and gastrojejunostomy were performed. Four dogs served as IORT only controls; one was unevaluable. Four dogs received WR2721, intraluminally at 720 mg/m2, in 16-18 ml Ringer's. Atraumatic clamps were placed on proximal and distal duodenum, without vascular compromise. WR2721 was injected into the duodenal lumen 30 minutes prior to IORT. Immediate postoperative recovery of the dogs receiving WR2721 was faster than controls. Necropsies were performed at 6 months. Grossly increased adhesions were noted in controls. Histopathologically, mucosal atrophy was greater in control dogs. Duodenal ulceration was noted in all controls, but in only one of four WR2721 dogs. Masson's trichrome and Verhoff Van Gieson stains demonstrated increased perivascular fibrosis, intimal proliferation, and fibrinoid medial necrosis of vessels in all controls, and one WR2721 dog. The other three WR2721 dogs had only mild perivascular fibrosis. Radioprotection, evaluated by the presence or absence of pancreatic atrophy, appeared to stop just beyond the bowel wall. In summary, WR2721 provided duodenal radioprotection in most dogs. The intraluminal administration of WR2721 allows decreased systemic side effects, and may eliminate tumor absorption. The study indicates that the intraluminal use of radioprotectors has broad potential application.

Hepatitis C infection in a pediatric dialysis population.

A variable prevalence of hepatitis C (HCV) infection has been reported in adult patients on hemodialysis. We have studied HCV infection and associated risk factors in a pediatric dialysis unit. Sera from all 27 patients undergoing either hemodialysis or peritoneal dialysis in our unit were tested for antibody to HCV by enzyme-linked immunosorbent assay, and seropositives were confirmed by recombinant immunoblot assay. Records were reviewed for demographic, biochemical, and risk factor data. From the total of 27 patients (12 male, mean age 20.9 years, range 7.3 to 28.1 years), five were anti-HCV(+) (18.5%). All the anti-HCV(+) patients had been on hemodialysis (69 to 194 months, mean 105 months), while of the 22 anti-HCV(-) patients, only 14 had been on hemodialysis (5 to 209 months, mean 41.4 months), P less than .005. All the anti-HCV(+) patients had received blood transfusions (10 to 124 units, mean 61.4 units) as had 12 of the anti-HCV(-) patients (1 to 54 units, mean 14 units), P less than .02. Of the 5 anti-HCV(+) patients, only one had prior hepatitis B infection; of the 22 anti-HCV(-) patients, three had hepatitis B surface antigen, and no others had evidence of hepatitis B infection. The most predictive risk factor for HCV infection was length of time on hemodialysis. Eleven of the 27 patients (40.7%) had abnormal alanine aminotransferase values, of whom four were anti-HCV(+), three were hepatitis B surface antigen(+), and one was seropositive for antibody to human immunodeficiency virus.(ABSTRACT TRUNCATED AT 250 WORDS)

Intraoperative radiation (IORT) injury to sciatic nerve in a large animal model.

Peripheral nerve appears to be a dose-limiting normal tissue in the clinical application of intraoperative radiation therapy (IORT). To assess IORT injury to peripheral nerve, three groups of five beagle dogs received doses of 12, 20 or 28 Gy to the surgically exposed and isolated right sciatic nerve in the mid-femoral region using 6 MeV electrons. The left sciatic nerve of each dog served as its own control. As a surgical control five dogs received surgical exposure of the nerve only. Monthly neurologic exams, electromyogram and nerve conduction studies were performed following treatment for 12 months. After that dogs were euthanatized and histologic studies of nerves were done to define the degree of axon and myelin loss as well as presence of fibrosis and vascular lesions for different doses of IORT. Results showed that the threshold dose most likely related to expression of severe radiation damage to the nerve in this model is between 20 and 25 Gy. Radiation injury to peripheral nerve appears to be the result of direct radiation effects on Schwann cells and nerve vasculature and secondary effects resulting from damage to regional muscle and vasculature. A theoretical mechanism of radiation injury to peripheral nerve is proposed.

The Xenopus clock gene is constitutively expressed in retinal photoreceptors.

Many aspects of normal retinal physiology are controlled by a retinal circadian clock. In Xenopus laevis, the photoreceptor cells within the retina contain a circadian clock that controls melatonin release. In this report we present the cloning and characterization of the Xenopus homolog of the Clock gene, known to be critical for normal circadian behavioral rhythms in the mouse. The Xenopus Clock gene is expressed primarily in photoreceptors within the eye and is expressed at constant levels throughout the day. Analysis of other tissues revealed that, as in other species, the Xenopus Clock gene is widely expressed. This characterization of the Clock gene provides a useful tool for further exploration of the role of the circadian clock in normal retinal function.

Effects of volume irradiated on the function of the canine ureter.

This study was designed to investigate the influence of the volume irradiated on the probability of ureteral complications and to provide data for volume modeling. One hundred thirty-four purpose-bred beagle dogs received single intraoperative doses of 6 MeV electrons ranging from 12 to 54 Gy to three lengths of ureter: 2, 4 or 8 cm. The response was evaluated by excretory urography. The ED50 was 21.9 Gy (95% CI 13.3-30 Gy) for 8 cm 3 years after treatment. The estimated ED50's were greater than 43 Gy for 4 cm and 85 Gy for 2 cm. Reducing the length of ureter irradiated from 8 cm to 4 cm increased the ED50 for ureteral dilation by at least a factor of 2, while reduction from 8 cm to 2 cm increased the ED50 by at least a factor of 4. The ED50 for renal injury secondary to stenosis was 30.5 Gy (95% CI 17.2-232 Gy) when an 8-cm field was irradiated. There was a significant effect of volume irradiated on the frequency of ureteral stenosis. Reducing the length of ureter included in the treatment field should allow delivery of higher doses to tumors without increased complications.