1 Treatments outcomes based on Frost and Hartl’s model are encouraging, but suggest that many sessions are required to produce change

and that clutter is slow to improve. The first case study reported that approximately 45 sessions were needed to completely reduce clutter.55 After 20 weeks of treatment, Steketee et al56 demonstrated a 16% reduction in Y-BOCS scores, while Saxena et al57 demonstrated a 35% reduction in Y-BOCS scores after 6 weeks of daily intensive treatment. Utilizing Steketee and Frost’s58 cognitive-behavioral treatment manual for compulsive hoarding, Tolin et al59 offered 26 individual sessions (in-office sessions and at least one home visit) over a 7- to 12-month period to 14 individuals. Inhibitors,research,lifescience,medical On average, treatment completers (n=10) demonstrated 25% improvement in their clutter and difficulty discarding, Inhibitors,research,lifescience,medical and 35% reduction in acquiring. Following this open trial, Steketee et al60 made minor modifications to the treatment and examined its efficacy in a randomized controlled trial. Findings from this trial indicated Inhibitors,research,lifescience,medical that improvements in hoarding symptoms were greater after receiving 12 sessions of cognitive behavioral therapy (CBT) than after waiting for a comparable period. After 26 sessions of CBT, 68% to 76% of patients were rated as improved by their therapists or themselves, respectively, and 41% of patients met criteria for clinically

significant improvement. Given that changes are slow to occur during

the treatment of compulsive hoarding, researchers have been examining alternative delivery models in hopes of increasing Inhibitors,research,lifescience,medical the cost-effectiveness of treatment. Using a multiple cohort pretest-post-test design, Muroff and colleagues examined Inhibitors,research,lifescience,medical the effectiveness of group CBT using Steketee and Frost’s treatment manual.32 After 16 to 20 sessions and two home visits, patients evidenced a mean reduction of 8.6 points on the Saving Inventory-Revised (SI-R), which is less than that produced from individual treatment using the same manual (18.7 or 16.9).59,60 After these investigators modified their research procedures to more thoroughly screen group members and utilized a more detailed and structured manual for the group, the Phosphoprotein phosphatase mean SI-R reduction in the final group was 14.25. As access to BEZ235 price clinicians trained in CBT for compulsive hoarding is limited, a Web-based self-help group has also been examined for its effectiveness. This Web-based treatment was also based on Steketee and Frost’s manual58 and required individuals to take active steps to reducing their hoarding behavior within 2 months of membership. After 6 months of memberships, SI-R scores decreased by an average of 6 points. These two group studies suggest that highly structured, in-person groups may lead to greater improvements in hoarding outcomes than less-structured groups.

Deficits in facial and acoustic expressions were found for posed and spontaneous expressions, suggesting a motor deficit. Moreover,

the impairments observed in these two channels correlate with each other and seem to be part, of broader deficits in expressiveness.5 They may reflect a deficit, in a premotor brain area involved in social and emotional expressions, such as the anterior cingulate area. The detrimental effect of deficits in expressiveness on social functioning and outcomes is an avenue of research. Although there is some evidence that impaired emotion expression in #SCH772984 keyword# schizophrenia has detrimental social consequences, this issue awaits confirmation. It is quite conceivable that deficits in expressiveness contribute to the stigma encountered by IWSs.42 Reactivity studies have brought contrasting results, and some studies have shown valence specificity. Overall, it can be concluded that emotion reactivity is not reduced in schizophrenia, and appears to be increased in specific conditions. Inhibitors,research,lifescience,medical Emotion experience Sixty-nine studies on emotion experience were reviewed. Emotion experience studies can be categorized according to the type of antecedents

(ie, emotional events) that they use: fixed stimulus in a laboratory setting Inhibitors,research,lifescience,medical or real-life emotion antecedents. In the first type of studies (evocative studies), the same emotional stimulus is presented to all participants, and they report, on their emotional experience after exposure to the stimulus. In the second type of studies (life-event studies), subjects are asked to evaluate events that happened during their lives. In life-event studies, two methodologies have been used: a time-sampling method and an event-sampling Inhibitors,research,lifescience,medical method. In the time-sampling method (often called “daily-life emotion studies”), emotional events are recorded over a defined time period. In the event-sampling method, subjects are asked to remember and describe past, events of a

specific emotional value (ic, when subjects felt the angriest or the happiest in their lives). The Inhibitors,research,lifescience,medical time-sampling method is most often prospective, whereas the event-sampling method is retrospective. Time-sampling studies give us access to events of moderate and low emotional intensity, whereas event-sampling studies allow us to examine antecedents of extreme emotional intensity. These two approaches first are therefore complementary. Besides these methodological issues, we will separately review two emotion phenomena: alexithymia and anhedonia. Recognition and awareness of own feelings (alexithymia) Impairments in identifying personal emotions have been described and identified in clinical groups, and they have been included in concepts like “alexithymia,” “emotion awareness,” and “emotional intelligence.” The most, widely used scale to measure alexithymia has been the Toronto Alexithymia Scale.

3 The cognitive syndrome is characterized by deficits in memory, language, visual constructional abilities, and other areas of intellectual functioning.4 The http://www.selleckchem.com/products/GDC-0449.html behavioral syndrome is characterized by symptoms of psychosis, aggression, depression, anxiety, agitation, and other common, but less well-defined, behavioral symptoms.5

Even though both syndromes have devastating consequences for patients and their care providers, it is the presence of behavioral symptoms and their impact on care providers that ultimately precipitate the chain of events that results in the demented patient’s placement in a. long-term care institution.6 This Inhibitors,research,lifescience,medical paper will focus on the behavioral and psychological symptoms Inhibitors,research,lifescience,medical of dementia. This focus was chosen because of the considerable treatment challenge these symptoms present, to clinicians and the burden they impose on patients, care providers, and society. Historical perspective Although physicians have been aware of the presence of behavioral symptoms in dementia since AD was first. described,7 a definition of these symptoms was not. attempted until 1986.8 In 1986, the syndrome defined was agitation, ie, “inappropriate verbal and motor behaviors that, are not, related to unmet needs or confusion per Inhibitors,research,lifescience,medical se.”9 Soon clinicians and researchers realized that the problem was more complex than the aforementioned

and that, patients presented not only symptoms of agitation, but also symptoms of aggression, psychosis, alteration in circadian rhythm, depression, and more.10 In 1996, the International Psychogeriatric Association

(IPA) called a consensus conference to examine the available knowledge on noncognitive symptoms of dementia. The goal of the conference was to achieve consensus Inhibitors,research,lifescience,medical on the use of more appropriate descriptive terms that would facilitate communication among researchers and therefore Inhibitors,research,lifescience,medical foster further development of the field. The experts in attendance agreed on an umbrella term that would include all behavioral symptoms observed in the dementia. The term was “Behavioral and Psychological Symptoms of Dementia” (BPSD),11 defined as “signs and symptoms of disturbed perception, thought content, mood or behavior that frequently occur in patients with dementia.”12 As new treatment options and scientific information emerged, another meeting was called by the IPA. This meeting evaluated the new data and concluded that a number of subsyndromes could be identified within the BPSD umbrella. These syndromes were Olopatadine psychosis, circadian rhythm (sleepwake) disturbance, depression, anxiety, agitation, and other less well-defined syndromes. The following is a brief summary of the current knowledge on BPSD with suggestions as to how this information can be applied to patient care. Etiology The etiology of BPSD is unknown. However, most experts probably agree that the etiology of BPSD is related to specific neuropathological brain lesions,13 psychological and environmental factors, or a. combination of both.

7 years, of the mediastinal hydatid cyst were reported from Iran.133-139 The symptoms related to the site of the pressure effect.139 Omentum and Retroperitoneum Seven cases of the mesenteric, diaphragmatic, omental, pelvic, and retroperitoneal hydatid cyst have been reported from Iran in the last 20 years.6,140-146 These cases may remain asymptomatic until reaching a large size,140 and the clinical signs vary according to the site. The parapharyngeal hydatid cyst in a 41-year-old female,147 and the nasolabial hydatid cyst

in an 11-year-old adolescent,148 were the last Inhibitors,research,lifescience,medical two extremely rare case reports in this review from Iran.147,148 Discussion Hydatid Roxadustat molecular weight disease is a unique parasitic disease that is endemic in many parts of the world.149 This parasitic disease is a significant public Inhibitors,research,lifescience,medical health concern in Iran, as an endemic country,150 rendering a review of the published cases of hydatid disease from this hyperendemic country vitally important. In hydatid disease, the liver and lung are the most

common Inhibitors,research,lifescience,medical involved organs, but the disease can be seen in any organ of the body.151 The rates of the localization of hydatid disease in different body organs vary in the literature.152 All the published cases in Iran included in this review are based on hospital experiences proven postoperatively by pathological examination. Our results demonstrated that the most common locations of the hydatid cyst, after the lung and liver, were the central nervous system, orbit, musculoskeletal system, cardiovascular system, kidney, and urinary tract. There were also reports of the spleen, uterus, Inhibitors,research,lifescience,medical ovary, pancreas, salivary gland, breast, adrenal, appendix, mediastinum, omentum, and retroperitoneum hydatid cysts. The clinical manifestations in the hydatid cyst of most parts of the body are too nonspecific to make a diagnosis based on the signs and symptoms Inhibitors,research,lifescience,medical before surgery.149-154 In all of the previous

reports from Iran and all around the world, it has been shown that serologic tests have many false-negative results, but imaging modalities such as ultrasonography, CT scan, and MRI have been the methods of choice, especially the latter, which has been the diagnostic method of choice for PAK6 the preoperative diagnosis of the hydatid cyst in most unusual locations.153 The best treatment for the hydatid cyst is surgical excision, accompanied by postoperative medical therapy.151 The next part of this review presents the salient points of each unusual site of the hydatid cyst extracted from the most recently published literature. Central Nervous System, Spinal Cord, and Orbit The cerebral and spinal cord hydatid cysts are very rare. Indeed, the existing literature contains about 300 articles,155 accounting for 2-3% of all the cases of hydatidosis.

The high prevalence rates suggest that any attempt to solve the pain crisis based upon increasing the availability of tertiary

care services is doomed, by weight of numbers, to fail. Thus the only viable solution to the crisis is empowerment of primary care physicians, especially family see more practitioners, who can be taught effective skills in chronic pain management. Inhibitors,research,lifescience,medical The model presented below is based upon empowering primary care physicians with education stressing theoretical knowledge together with practical hands-on clinically oriented learning. The model constitutes a three-tiered pyramid, each tier narrower than the one below, from the lower-tiered primary care physicians who should be trained

as pain trustees, up to the second-tiered secondary care or community consultant physicians, and up to the tertiary center-based specialists in pain medicine ( Figure 1 ). In our model, the vast majority of pain patients should be treated in the primary care setting. Inhibitors,research,lifescience,medical Patients suffering from more complex problems should be referred to secondary, Inhibitors,research,lifescience,medical community-based consultants, probably working within a small multidisciplinary setup. These consultants could either be pain specialist working in the community or primary care physicians who have undergone further training and are pain trustees with an a additional diploma in pain Inhibitors,research,lifescience,medical and musculoskeletal medicine. Finally, the top tier should be populated by specialists in pain medicine who have undergone extensive further training, especially in the utilization of invasive procedures. Only the most complex of cases, and those needing specialized care, should be referred to tertiary pain centers. Figure 1 The Pyramid Model for the Stratification of Chronic Pain Treatment in the Community. The main challenge in realizing this model lies in the training of doctors according to these

tiers, empowering them with knowledge Inhibitors,research,lifescience,medical and skills necessary for the task. There are already a few year-long courses being taught in Israel, most notably in the Rambam School of Pain Medicine in association with the Technion, at Tel Aviv University in association with the family practice association, and at the Beer Sheva School of Medicine. In the following section we will give a detailed description of the necessary training much of the various levels (see Figure 1 and Table 1 ). Table 1 Operative Plan for Implementing the Pyramid Model. PRIMARY CARE PHYSICIANS Patients who suffer chronic pain are primarily seen by family physicians and, to a lesser extent, by orthopedic surgeons, neurologists, rheumatologists, and other specialists. Since most of these physicians have not received sufficient training in the treatment of pain as students and as residents, their knowledge in the field is based primarily on postgraduate education.

Distinction of this from the regional heterogeneity of dilated cardiomyopathy may be supported by ischemic or viable responses to either exercise or pharmacologic stress echocardiography,9) as well as coronary imaging with CT and scar imaging with CMR. Likewise, ambiguity about the cause of LV thickening in

hypertensive heart failure may be elucidated #VX-770 datasheet keyword# by techniques characterizing myocardial infiltration. Finally, while apical ballooning or mid to basal ballooning can be a clue of stress-induced cardiomyopathy (SCMP),10),11) other testing may be needed to exclude acute myocardial infarction (AMI). Role of CMR Although echocardiography is extremely versatile and readily accessible, image quality is often limited, and geometric assumptions are required to quantify LV systolic function. Inhibitors,research,lifescience,medical Furthermore, it lacks the ability to provide

more detailed tissue characterization, which can be extremely important in defining the etiology of heart failure. CMR is currently considered the gold standard for the assessment of LV mass, systolic function, and assessment of myocardial fibrosis. CMR has the ability to image in any three dimensional plane, offering the ability to produce extremely accurate and reproducible assessment of LV and right ventricular (RV) volumes, ejection fraction, Inhibitors,research,lifescience,medical and mass, without relying on geometric assumptions that can result in significant miscalculations particularly in dilated ventricles.12),13) In addition, CMR offers the ability Inhibitors,research,lifescience,medical to assess myocardial perfusion as well as implementing other imaging techniques [delayed hyperenhancement (DHE) imaging, T1-weighted, T2-weighted, and fat suppression imaging techniques] to assess for myocardial fibrosis as well as myocardial edema. DHE imaging allows for the identification of myocardial fibrosis with high resolution and offers the ability to differentiate between types of cardiomyopathies, based on patterns of fibrosis (Fig. 1).14),15) DHE-CMR can identify Inhibitors,research,lifescience,medical significant coronary artery disease and

decrease the need of conventional coronary angiography in patients presenting with heart failure of uncertain etiology.16) T1- or T2-weighted image sequences provide the ability to differentiate between fat, muscle, and areas of inflammation, Casein kinase 1 based on the different proton relaxation properties of these tissues. Tissue edema appears bright on T2-weighted images in both acute coronary syndromes as well as inflammatory processes such as cardiac sarcoidosis or myocarditis.17),18) Myocardial edema may occur in isolation, but is often accompanied by characteristic patterns of myocardial fibrosis, which has the ability to elucidate the etiology of decreased ventricular function. The standard T2-weighted image sequences use turbo spin-echo sequences, and have been limited by artifacts (e.g., posterior wall signal loss caused by through-plane motion or bright rim artifacts caused by stagnant blood along the endocardial surface).

Table 2 Distribution of the reasons to entry the ED recorded by using ICPC 2 classification in group E patients (N = 1244) of the Espoo EDs Discussion The implementation

of the ABCDE-triage combined with public guidance was associated with reduction in the number of patient visits to GP out-of-hours ED services by about 24%. The observed Inhibitors,research,lifescience,medical reduction in GP visits in the ED may partly be due to considerable public debate and the publicity provided by the new system and rules. It is possible that some of the patients decided not to request emergency care at all due to the expected long waiting times or risk of being redirected to daytime health services. Patients were Inhibitors,research,lifescience,medical also assessed to group E by the triage nurse and redirected to homecare. This result is higher than our former experience from Vantaa City where the number of ED visits decreased by 8% after implementation of ABCDE-triage [16]. In Espoo, the population seemed to adapt very quickly to the idea that those who needed help most must go first and those whose

need is not urgent should not selleck kinase inhibitor necessarily visit the ED at all. However, a considerable difference between Vantaa and Espoo was, that in Espoo the patient who was assessed to group E might be sent home with advice Inhibitors,research,lifescience,medical while in Vantaa the patient was allowed to stay and wait as long as the queue of more urgent patients (groups A-D) persisted [16]. This may also have explained why the decrease in patient visits was much higher in Espoo than in Vantaa. GPs were previously assumed to regulate access to the acute secondary health care by referring those patients who need specialist Inhibitors,research,lifescience,medical care. The triage was performed by primary health care in EDs but it did not diminish or increase the workload of the secondary health care in the same facility. Altogether, the present finding agrees with the former Inhibitors,research,lifescience,medical report of Vertesi [3] which suggested that triage did not automatically enhance activities in the secondary health care ED. The number of visits to primary care GPs during office hours was unchanged from March 2004

to February 2008 in Espoo (Figure ​(Figure3).3). Thus, the decrease in patient visits to the GPs in Espoo EDs did not cause an overflow of patients in the office hour GP practice. There were some hints that demand for nurse visits in daytime services increased but this could not be verified because also other changes were made in office-hour Suplatast tosilate public health to alter the workload of nurses at the same time. Furthermore, no excessive doctor resources were allocated to office-hour activities at the time of the intervention. Thus, we cannot exclude the possibility that the lack of change in the number of visits to primary care GPs during office hours was just attributable to that fact. Yet the same phenomenon was observed in Vantaa City in our previous work [16].

This may represent the preservation of a strategy that the subjects developed in the first study to solve the neurocognitive task. For example, we found that stereotypy, a component cognitive process/strategy based on repetitive sequential

selection of a correct spatial pattern sequence, was preserved in subjects after MST, and resulted in their increased accuracy on a spatial working memory task.103 However, when administered ECS, subjects Inhibitors,research,lifescience,medical showed decreased stereotypy and poorer accuracy on the same spatial working memory measure. Thus, MST allows for the maintenance of component cognitive processes/strategies, which are integral to higher order cognitive functions.

Indeed, neuropsychologic research with healthy human subjects has found strong associations between cognitive strategies and neurocognitive outcome. Importantly, on commonly used neuropsychologic measures (eg, digit span, category fluency), strategy was more strongly associated Inhibitors,research,lifescience,medical with performance than were other demographic factors such as age and education.106 Antidepressant activity of magnetic seizure therapy Although still in developmental phases, controlled clinical case reports and series, and Inhibitors,research,lifescience,medical one randomized control trial have found MST to have antidepressant benefits in patients with MDD or bipolar disorder. Four case reports that varied in methodologic design through use of various coil types (eg, round, butterfly, figure-of-eight), dosing parameters (eg, between 50 Hz and 100 Hz stimulation; 200 to 1000 pulses) and administration of number of treatment sessions (eg, four to twelve) Inhibitors,research,lifescience,medical found patients maintained global cognitive abilities, had rapid rates of reorientation recovery, and

had corresponding decreased depression severity.85,107-109 Inhibitors,research,lifescience,medical Two case series showed similar results in patients with MDD.110,111 In the first, only two MST sessions (provided up to 60 Hz with multiple coil types) were administered Oxygenase in the context of an acute ECT course.110 Both treatments resulted in similar decreases in depression severity, but only in the MST sessions did patients show quick orientation recovery and preservation of multiple cognitive abilities including processing speed, and simple attention and memory. In the second case series, which followed a similar design as the first, 11 patients received at least one MST session (provided at 100 Hz for 10 seconds with a round coil).111 The study found that relative to ECT, patients showed quicker recovery of orientation that was shorter on RGFP966 molecular weight average by approximately 15 minutes. To date, a randomized controlled trial found MST (delivered with twin coil, 100 Hz, up to 600 pulses) and ECT (delivered with 0.

Induction of diabetes decreased testicular StAR mRNA expression by 66% and MAE treatment enhanced mRNA expression to the same level of the control group. However, the expression of P540scc was not significantly decreased in the diabetic group as compared to the control group. Conclusion: Our findings indicated that MAE significantly increased Ts production in the diabetic rats, probably through the induction of StAR mRNA expression levels. Administration of MAE to

experimental models of diabetes can effectively Inhibitors,research,lifescience,medical attenuate oxidative stress-mediated testosterone depletion. Keywords: Diabetes mellitus, Morus alba, Oxidative stress, Testosterone Introduction Diabetes mellitus (DM) Inhibitors,research,lifescience,medical is one of the most common chronic diseases worldwide. It has been estimated that the prevalence of DM will increase from 275 million adults in 2010 to 439 million by 2030.1 It has been posited that oxidative stress plays a pivotal role in the pathogeneses of DM.2 Accumulated data suggest that DM

is linked with male reproductive dysfunction.3 In rats, diabetes induces apoptosis in the testicular germ cell4 and decreases sperm count and plasma testosterone (Ts) levels.5 However, the potential contribution Inhibitors,research,lifescience,medical of oxidative stress due to diabetic condition in the development of testicular abnormalities has not been fully clarified. It has been shown that reactive oxygen species (ROS) can inhibit the steroid hormone production of cultured leydig cells by directly Inhibitors,research,lifescience,medical affecting the steroidogenic enzymes.6 Be that as it may, the testicular

activity of steroidogenic proteins under diabetic condition has yet to be studied. Steroid acute regulatory protein (StAR) and P450 cholesterol side-chain cleavage enzyme (P450scc) are two important proteins that catalyze the first steps in steroidogenesis.7 Diemer et al.8 demonstrated that the in vitro exposure of MA-10 tumor leydig cells to ROS decreased StAR protein expression levels. Morus alba (mulberry) is a plant rich in phytochemicals, which have an important role in diet-based therapies to cure several diseases.9 Antioxidant and antidiabetic activity of Morus alba leaf extract (MAE) in Streptozotocin-induced Inhibitors,research,lifescience,medical diabetic rats has been shown.10,11 The objective of this study was to investigate the effect of the long-term administration of MAE on oxidative stress markers and steroidogenesis in diabetic rats. The likely mechanism of Digestive enzyme MAE action on steroidogenesis was also explored. Materials and Methods Chemicals Streptozotocin, thiobarbituric acid (TBA), 1,1,3,3-tetramethoxypropane, reduced glutathione (GSH), oxidized glutathione (GSSG), nicotine-amid-adenine-dinucleotide phosphate (NADPH), glutathione reductase (GR), tripyridyl-s-triazine (TPTZ), and other Ibrutinib nmr high-grade chemicals were purchased from Sigma Chemical Company (St. Louis, Missouri, USA). Preparation of Morus Alba Leaf Extract Leaves of Morus alba were collected from a local area (Shiraz, Iran) in April.

GUSB enzyme activity was increased >10-fold in brain, liver, spleen, lung, and kidney, but not in heart (Figure 3(b)). The expression pattern of GUSB gene among mouse

organs in vivo is consistent with the local expression of the TfR in the vascular barriers of these tissues. The liver and spleen are perfused with fenestrated capillaries that are highly porous, so the 100nm THLs can freely cross their vascular barrier [20]. Heart, lung, and kidney are perfused with capillaries with continuous endothelial barriers [38]. Thus, the observation that GUSB enzyme activity is increased in lung and kidney with TfRMAb-targeted THLs in vivo provides additional evidence for Inhibitors,research,lifescience,medical the expression of the TfR on the vascular barrier in these organs in the mouse [27]. Inhibitors,research,lifescience,medical Lack of expression in heart supports prior work with reporter genes showing that TfRMAb-targeted THLs are not delivered across the vascular barrier in heart [20, 21, 27]. The brain GUSB enzyme activity observed at 48h after a single THL administration approximated

2U/mg protein (Figure 3(b)), which represents 55% of the brain level in heterozygotes [39]. Since the replacement of just 1–5% of lysosomal enzyme activity in an organ may Inhibitors,research,lifescience,medical be sufficient to cause therapeutic AZD0530 supplier effects and a reversal of lysosomal storage disease [37], the levels of GUSB enzyme activity generated in the brain of null mice with a single Inhibitors,research,lifescience,medical IV injection of THLs is within the therapeutic range. The plasmid DNA is expressed episomally in brain cells without integration into the host genome [33]. Therefore, long-term treatment of lysosomal storage disorders with intravenous administration of THLs will require repeat administration of the gene medicine at intervals that are determined by both the persistence of transgene expression and the turnover of the expressed protein in brain and peripheral organs. 5. Brain Expression of Therapeutic

Genes in a Model of Parkinson’s Disease The therapeutic efficacy of THLs was demonstrated in vivo in a model of Inhibitors,research,lifescience,medical Parkinson’s disease (PD), wherein the therapeutic gene encoded for tyrosine hydroxylase (TH) [30]. PD is associated with a loss of dopaminergic neurons in the substantia nigra, which terminate in the striatum [40, 41]. The rate limiting enzyme in the synthesis of dopamine is TH, and a potential treatment for PD is TH gene replacement therapy. Studies were performed in the rat 6-hydroxydopamine Etomidate (6-OHDA) model, and with THL packaged with a TH expression plasmid driven by the Gfap brain-specific promoter, designated clone 951 [30]. Gfap-TH-THLs were constructed with the OX26 MAb to target the rat TfR (Table 1). The intracerebral injection of 6-OHDA produced a 98% reduction in the levels of TH in the ipsilateral striatum as compared with the contralateral or nonlesioned control animals (Table 2).