The TNFR1 antagonist Atrosimab reduces neuronal loss, glial activation and memory deficits in an acute mouse model of neurodegeneration.

Tumor necrosis factor alpha (TNF-α) and its key role in modulating immune responses has been widely recognized as a therapeutic target for inflammatory and neurodegenerative diseases. Even though inhibition of TNF-α is beneficial for the treatment of certain inflammatory diseases, total neutralization of TNF-α largely failed in the treatment of neurodegenerative diseases. TNF-α exerts distinct functions depending on interaction with its two TNF receptors, whereby TNF receptor 1 (TNFR1) is associated with neuroinflammation and apoptosis and TNF receptor 2 (TNFR2) with neuroprotection and immune regulation. Here, we investigated the effect of administering the TNFR1-specific antagonist Atrosimab, as strategy to block TNFR1 signaling while maintaining TNFR2 signaling unaltered, in an acute mouse model for neurodegeneration. In this model, a NMDA-induced lesion that mimics various hallmarks of neurodegenerative diseases, such as memory loss and cell death, was created in the nucleus basalis magnocellularis and Atrosimab or control protein was administered centrally. We showed that Atrosimab attenuated cognitive impairments and reduced neuroinflammation and neuronal cell death. Our results demonstrate that Atrosimab is effective in ameliorating disease symptoms in an acute neurodegenerative mouse model. Altogether, our study indicates that Atrosimab may be a promising candidate for the development of a therapeutic strategy for the treatment of neurodegenerative diseases.

The effects of forensic mental health reports on decisions about guilt in the Netherlands: An experimental approach.

In the Netherlands, in approximately 30% of the more serious criminal cases, a pretrial forensic mental health report (FMHR) is requested to inform the court whether a mental disorder was present at the time of alleged crime, whether this disorder affected behavior and decision-making at the time of the offense and how this disorder may affect future (criminal) behavior. While informative for sentencing decisions, information about mental disorders or risk is irrelevant for the question whether the defendant committed the alleged crime. Yet based on cognitive psychological theory of evidence evaluation and integration, we hypothesized that information in an FMHR would affect the evaluation of evidence as well as the ultimate decision about guilt. Using an experimental vignette study among 200 law and criminology students with manipulation of the presence and content of an FMHR, we found a main effect of the presence of an FMHR report on decisions about guilt. The proportion of guilty verdicts increased with almost 20% when an FMHR was present compared to when this report was absent, irrespective of the type of disorder (schizophrenia or personality disorder) or level of recidivism risk (low or high) present in the report. We did not find support for our hypothesis that this effect could be explained by assimilation of other available evidence. Implications for further research and practice are discussed.

Lipocalin 2 contributes to brain iron dysregulation but does not affect cognition, plaque load, and glial activation in the J20 Alzheimer mouse model.

BACKGROUND: Lipocalin 2 (Lcn2) is an acute-phase protein implicated in multiple neurodegenerative conditions. Interestingly, both neuroprotective and neurodegenerative effects have been described for Lcn2. Increased Lcn2 levels were found in human post-mortem Alzheimer (AD) brain tissue, and in vitro studies indicated that Lcn2 aggravates amyloid-ß-induced toxicity. However, the role of Lcn2 has not been studied in an in vivo AD model. Therefore, in the current study, the effects of Lcn2 were studied in the J20 mouse model of AD. METHODS: J20 mice and Lcn2-deficient J20 (J20xLcn2 KO) mice were compared at the behavioral and neuropathological level. RESULTS: J20xLcn2 KO and J20 mice presented equally strong AD-like behavioral changes, cognitive impairment, plaque load, and glial activation. Interestingly, hippocampal iron accumulation was significantly decreased in J20xLcn2 KO mice as compared to J20 mice. CONCLUSIONS: Lcn2 contributes to AD-like brain iron dysregulation, and future research should further explore the importance of Lcn2 in AD.

Short-Term Effects of Imprisonment Length on Recidivism in the Netherlands.

This article assesses the relationship between imprisonment length and recidivism. The data come from a unique longitudinal and nationwide study of Dutch prisoners, serving an average of 4.1 months of confinement (N = 1,467). A propensity score methodology is used to examine the dose-response relationship for three types of registered recidivism (i.e., reoffending, reconviction, and reincarceration) within a 6-month follow-up period. Findings indicate that length of imprisonment exerts an overall null effect on future rates of recidivism and that this conclusion holds across the various types of recidivism. These findings contribute to continuing scholarly debates over the social and economic costs of imprisonment.

WHOLE BODY VIBRATION IMPROVES ATTENTION AND MOTOR PERFORMANCE IN MICE DEPENDING ON THE DURATION OF THE WHOLE-BODY VIBRATION SESSION.

BACKGROUND: Whole body vibration (WBV) is a form of physical stimulation via mechanical vibrations transmitted to a subject. It is assumed that WBV induces sensory stimulation in cortical brain regions through the activation of skin and muscle receptors responding to the vibration. The effects of WBV on muscle strength are well described. However, little is known about the impact of WBV on the brain. Recently, it was shown in humans that WBV improves attention in an acute WBV protocol. Preclinical research is needed to unravel the underlying brain mechanism. As a first step, we examined whether chronic WBV improves attention in mice. MATERIAL AND METHODS: A custom made vibrating platform for mice with low intensity vibrations was used. Male CD1 mice (3 months of age) received five weeks WBV (30 Hz; 1.9 G), five days a week with sessions of five (n=12) or 30 (n=10) minutes. Control mice (pseudo-WBV; n=12 and 10 for the five and 30 minute sessions, respectively) were treated in a similar way, but did not receive the actual vibration. Object recognition tasks were used as an attention test (novel and spatial object recognition - the primary outcome measure). A Balance beam was used for motor performance, serving as a secondary outcome measure. RESULTS: WBV sessions of five (but not WBV sessions of 30 minutes) improved balance beam performance (mice gained 28% in time needed to cross the beam) and novel object recognition (mice paid significantly more attention to the novel object) as compared to pseudo WBV, but no change was found for spatial object performance (mice did not notice the relocation). Although 30 minutes WBV sessions were not beneficial, it did not impair either attention or motor performance. CONCLUSION: These results show that brief sessions of WBV improve, next to motor performance, attention for object recognition, but not spatial cues of the objects. The selective improvement of attention in mice opens the avenue to unravel the underlying brain mechanisms.

Differential reporting of mixed DNA profiles and its impact on jurists' evaluation of evidence. An international analysis.

While DNA analysis is considered by many the gold standard in forensic science, there is ample room for variation in interpretation and reporting. This seems especially the case when working with (complex) mixed DNA profiles. Two consecutive studies on differential DNA reporting were conducted. In Study 1, we first examined type and magnitude of differences when forensic DNA experts across institutes and jurisdictions are handed an identical forensic case with mixed profiles. In Study 2, we explore the impact of the observed differential reporting on jurists' evaluation of the DNA evidence. 19 DNA expert reports from forensic institutes across Western jurisdictions were obtained. Differences between the reports were many and include extensiveness of the reports, explanations of technical issues, use of explanatory appendices, level of reporting, use of context information, and, most markedly, type and substantive content of the conclusions. In Study 2, a group of criminal law students judged a selection of these reports in a quasi experimental study design. Findings show that these differing reports have quite different evidentiary value for jurists, depending on which expert authored the report. It is argued that the impact of differential reporting on jurists' evaluation was so fundamental and substantive that it is seems reasonable to claim that in an actual court case it could make the difference between acquittal and conviction.

Are stalkers recidivists? Repeated offending by convicted stalkers.

Stalking is an obsessive behavior. Legal definitions generally characterize stalking as repetitive conduct. It may therefore be expected that recidivism by stalkers is high. We investigated court statistics of stalking cases to establish which proportion relapses in stalking behavior after a conviction and what other types of new crimes they commit. Case files of stalking cases have been investigated to find out whether and which neutralization techniques are used by stalkers to justify harassing behaviors. Stalkers who do recidivate do so quickly after a conviction. They appear to make use of various neutralization techniques. There is a small group of highly obsessive stalkers that seems not to be stopped by any of the measures, sanctions, or interventions that are imposed.

Sleep deprivation impairs spatial working memory and reduces hippocampal AMPA receptor phosphorylation.

Sleep is important for brain function and cognitive performance. Sleep deprivation (SD) may affect subsequent learning capacity and ability to form new memories, particularly in the case of hippocampus-dependent tasks. In the present study we examined whether SD for 6 or 12 h during the normal resting phase prior to learning affects hippocampus-dependent working memory in mice. In addition, we determined effects of SD on hippocampal glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors and their regulatory pathways, which are crucially involved in working memory. After 12 h SD, but not yet after 6 h, spatial working memory in a novel arm recognition task was significantly impaired. This deficit was not likely due to stress as corticosterone levels after SD were not significantly different between groups. In parallel with the change in cognitive function, we found that 12 h SD significantly reduced hippocampal AMPA receptor phosphorylation at the GluR1-S845 site, which is important for incorporation of the receptors into the membrane. SD did not affect protein levels of cyclic-AMP-dependent protein kinase A (PKA) or phosphatase calcineurin (CaN), which regulate GluR1 phosphorylation. However, SD did reduce the expression of the scaffolding molecule A-kinase anchoring protein 150 (AKAP150), which binds and partly controls the actions of PKA and CaN. In conclusion, a relatively short SD during the normal resting phase may affect spatial working memory in mice by reducing hippocampal AMPA receptor function through a change in AKAP150 levels. Together, these findings provide further insight into the possible mechanism of SD-induced hippocampal dysfunction and memory impairment.

Neuroprotective effects of vinpocetine and its major metabolite cis-apovincaminic acid on NMDA-induced neurotoxicity in a rat entorhinal cortex lesion model.

Vinpocetine (ethyl-apovincaminate, Cavinton), a synthetic derivative of the Vinca minor alkaloid vincamine, has been used now for decades for prevention and treatment of cerebrovascular diseases predisposing to development of dementia. Both vinpocetine and its main metabolite cis-apovincaminic acid (cAVA) exert a neuroprotective type of action. Bilateral N-methyl-D-aspartate (NMDA)-induced neurodegeneration in the entorhinal cortex of rat was used as a dementia model to confirm the neuroprotective action of these compounds in vivo. NMDA-lesioned rats were treated 60 min before lesion and throughout 3 postoperative days with a 10 mg/kg intraperitoneal dose of vinpocetine or cAVA. Behavioral tests started after termination of drug treatment and consisted of novel object recognition, social discrimination, and spontaneous alternation in a Y-maze, and spatial learning in the Morris water maze. At the end of behavioral testing brains were perfused with fixative and the size of the excitotoxic neuronal lesion and that of microglial activation around the lesion were assayed quantitatively on brain sections immunostained for neuron-specific nuclear protein (NeuN) and integrin CD11b, respectively. Entorhinal NMDA lesions impaired recognition of novel objects and the new social partner, and suppressed spontaneous alternation and spatial learning performance in the Morris maze. Both vinpocetine and cAVA effectively attenuated the behavioral deficits, and significantly decreased lesion size and the region of microglia activation. Both lesion-induced attention deficit and learning disabilities were markedly alleviated by vinpocetine and cAVA. The morphological findings corroborated the behavioral observations and indicated reduced lesion size and microglia activation especially after vinpocetine treatment which supports an in vivo neuroprotective mode of action of vinpocitine and a less potent action of cAVA.

Repetitive stimulation of adenosine A1 receptors in vivo: changes in receptor numbers, G-proteins and A1 receptor agonist-induced hypothermia.

Adenosine is an important neuromodulator and neuroprotective molecule, which is produced in the brain as a function of neuronal activity, coupling energy expenditure to energy supply. Under conditions of increased need and reduced availability of energy, including hypoxia and prolonged wakefulness, there is an increase in adenosine turnover and adenosine receptor stimulation. The aim of the present study was to examine how repetitive adenosine receptor stimulation affects receptor function and adenosinergic signaling in the brain. Adult male Wistar rats received daily intraperitoneal injections of the adenosine A1 receptor agonist N(6)-cyclopentyladenosine (CPA; 0.25 mg/kg; once per day) and effects on adenosine signaling were established with receptor and G-protein autoradiography. Injections of CPA for 5 consecutive days caused a significant decrease in adenosine A1 receptor numbers in the hippocampus and somatosensory cortex. In contrast, while the amount of adenosine A1 receptor-activated G-proteins was not affected in most regions, a significant increase was found in the somatosensory cortex. On the level of physiological output, CPA-induced hypothermia was significantly attenuated, suggesting a functional desensitization of the A1 receptor system. Taken together, the present findings suggest that repetitive stimulation of the A1 receptors can affect elements of the adenosinergic signaling cascade in the rat brain in a region-specific manner.

Aging is accompanied by a subfield-specific reduction of serotonergic fibers in the tree shrew hippocampal formation.

The hippocampal formation is a crucial structure for learning and memory, and serotonin together with other neurotransmitters is essential in these processes. Although the effects of aging on various neurotransmitter systems in the hippocampus have been extensively investigated, it is not entirely clear whether or how the hippocampal serotonergic innervation changes during aging. Rat studies, which have mostly focused on aging-related changes in the dentate gyrus, have implied a loss of hippocampal serotonergic fibers. We used the tree shrew (Tupaia belangeri), an intermediate between insectivores and primates, as a model of aging. We applied immunocytochemistry with an antibody against serotonin to assess serotonergic fiber densities in the various hippocampal subfields of adult (0.9-1.3 years) and old (5-7 years) tree shrews. Our results have revealed a reduction of serotonergic fiber densities in the stratum radiatum of CA1 and CA3, and in the stratum oriens of CA3. A partial depletion of serotonin in the hippocampal formation, as can be expected from our current observations, will probably have an impact on the functioning of hippocampal principal neurons. Our findings also indicate that the rat and the tree shrew hippocampal serotonergic innervation show some variations that seem to be differentially affected during aging.

Input from the medial septum regulates adult hippocampal neurogenesis.

Neural progenitors in the subgranular zone of the hippocampal formation form a continuously proliferating cell population, generating new granule neurons throughout adult life. Between 10 days and 1 month after their formation, many of the newly generated cells die. The present study investigated whether a partial lesion of one of the main nuclei projecting to the hippocampus, the medial septum (MS), affects survival and differentiation of cells during this critical period. Rats were injected with BrdU and 5 days later excitotoxic lesion of the MS was applied by infusion of either 30 or 60 nmol of N-methyl-D-aspartate (NMDA). One week after the lesion, quantification of immunopositive cells revealed that the number of GABAergic cells was significantly reduced in both lesioned groups, whereas a decline in cholinergic cell number was observed only after injection of 60 nmol of NMDA. The partial septohippocampal denervation significantly reduced hippocampal neurogenesis. Survival of newly generated neurons was decreased by approximately 40%. The MS lesion did not affect proliferation of hippocampal progenitors. The present study points out the importance of a functional septohippocampal pathway for the regulation of hippocampal neurogenesis and highlights the potential role of GABA as a mediator in this phenomenon.

Galantamine-induced behavioral recovery after sublethal excitotoxic lesions to the rat medial septum.

Clinical trials show beneficial effects of acetylcholinesterase (AChE) inhibitors, including galantamine, on cognitive functions in patients with mild to moderate Alzheimer's disease. Galantamine shows a dual action profile by also acting as an allosteric modulator of nicotinic acetylcholine receptors. Nevertheless, its in vivo mechanism of action is only partly understood. Here, we first established a novel lesion model provoking significant functional impairment of the septo-hippocampal projection system without triggering massive neuronal death in the rat medial septum. Next, we studied whether galantamine, administered in doses of 1 and 3mg/kg post-lesion, promotes functional recovery of spatial navigation behaviors, and affects the output of septal cholinergic projections. Infusion of N-methyl-d-aspartate (NMDA; 30nmol/1microl) in the medial septum resulted in spatial learning deficits associated with significant shrinkage of cholinergic neurons and reduced AChE activity in the hippocampus at 7 days post-lesion. Galantamine treatment alone significantly increased the hippocampal acetylcholine concentration and attenuated the NMDA-induced spatial learning impairment. Galantamine post-treatment also affected NMDA-induced changes in AChE and choline-acetyltransferase activities. In conclusion, our data show that galantamine attenuates experimentally-induced cognitive impairments underscored by mild neuronal damage.

Ageing-related decline in adenosine A1 receptor binding in the rat brain: an autoradiographic study.

The adenosine system has important neuromodulatory and neuroprotective functions in the brain. Several lines of evidence suggest that ageing is associated with major alterations in the adenosine system, which may be partially responsible for changes in sleep, mood, and cognition. In the present study, we examined adenosine A1 receptor density in the rat brain by means of quantitative autoradiography to obtain a detailed anatomical overview of the changes during ageing. A1 receptor binding was assessed in young, old, and senescent animals of 3, 24, and 30 months old, respectively. There was a clear age-dependent reduction in adenosine A1 receptors in most of the brain areas examined, but the magnitude of this reduction varied greatly among regions. Also, whereas some regions displayed a gradual decline in A1 binding sites across the three age classes, other regions showed a particularly strong decrease between the ages of 24 and 30 months. For example, whereas the hippocampus and thalamus showed a gradual decline in A1 binding, some cortical and septal regions showed a more abrupt decline after the age of 24 months. Since particularly in rats many studies have used animals at the age of 24 months or even less, the ageing-related decline in adenosine A1 signaling might have been underestimated.

Comparison of 2 commercially available chlorhexidine mouthrinses.

BACKGROUND: It has been shown that lowering the concentration of chlorhexidine (CHX) in mouthrinses from 0.2% to 0.12% does not adversely affect inhibition of plaque accumulation. The aim of this study was to evaluate the inhibition of plaque growth by 2 commercially available mouthrinses used according to the manufacturers' instructions, one with a 30-second and one with a 60-second rinsing time. METHODS: The clinical investigation was a single-blind, randomized study involving 80 volunteers (40 female, 40 male; mean age 25.7 years). At the start of the trial, all participants received a dental prophylaxis to remove all plaque deposits. Subjects refrained from all mechanical oral hygiene procedures, but rinsed twice a day with the allocated CHX mouthrinse over a period of 72 hours. The chlorhexidine preparations compared were a 0.12% concentration used at a 15 ml dose for a rinsing time of 30 seconds and a 0.2% concentration used at a dose of 10 ml for 60 seconds. After 72 hours, the plaque index (PI) from all volunteers was recorded at 6 sites per tooth. All participants received a questionnaire to evaluate their perception of the mouthrinses. RESULTS: After 72 hours, the 15 ml/30 second/0.12% CHX group had a mean whole mouth PI of 1.65 (SD 0.31) compared with a mean PI of 1.60 (SD 0.40) for the 10 ml/60 second/0.2% CHX group. The difference in plaque scores between the groups was not statistically significant. Results from the questionnaire showed no significant difference between the groups for taste perception, duration of taste, alteration in taste, or perceived plaque reduction; however, the panelists preferred the shorter rinsing time of 30 seconds and, for this parameter, the difference was statistically significant (P = 0.048). CONCLUSIONS: The results of this short-term study showed that there was no statistically significant difference between both commercially available CHX mouthrinses with respect to plaque inhibition, although both differed in concentration and rinsing time. The subject preference phase of the study indicated that the shorter rinsing time of 30 seconds was favored.

Rabbit forebrain cholinergic system: morphological characterization of nuclei and distribution of cholinergic terminals in the cerebral cortex and hippocampus.

Although the rabbit brain, in particular the basal forebrain cholinergic system, has become a common model for neuropathological changes associated with Alzheimer's disease, detailed neuroanatomical studies on the morphological organization of basal forebrain cholinergic nuclei and on their output pathways are still awaited. Therefore, we performed quantitative choline acetyltransferase (ChAT) immunocytochemistry to localize major cholinergic nuclei and to determine the number of respective cholinergic neurons in the rabbit forebrain. The density of ChAT-immunoreactive terminals in layer V of distinct neocortical territories and in hippocampal subfields was also measured. Another cholinergic marker, the low-affinity neurotrophin receptor (p75(NTR)), was also employed to identify subsets of cholinergic neurons. Double-immunofluorescence labeling of ChAT and p75(NTR), calbindin D-28k (CB), parvalbumin, calretinin, neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase, or substance P was used to elucidate the neuroanatomical borders of cholinergic nuclei and to analyze the neurochemical complexity of cholinergic cell populations. Cholinergic projection neurons with heterogeneous densities were found in the medial septum, vertical and horizontal diagonal bands of Broca, ventral pallidum, and magnocellular nucleus basalis (MBN)/substantia innominata (SI) complex; cholinergic interneurons were observed in the caudate nucleus, putamen, accumbens nucleus, and olfactory tubercule, whereas the globus pallidus was devoid of cholinergic nerve cells. Cholinergic interneurons were frequently present in the hippocampus and to a lesser extent in cerebral cortex. Cholinergic projection neurons, except those localized in SI, abundantly expressed p75(NTR), and a subset of cholinergic neurons in posterior MBN was immunoreactive for CB and nNOS. A strict laminar distribution pattern of cholinergic terminals was recorded both in the cerebral cortex and in CA1-CA3 and dentate gyrus of the hippocampus. In summary, the structural organization and chemoarchitecture of rabbit basal forebrain may be considered as a transition between that of rodents and that of primates.

Explaining capital punishment support in an abolitionist country: the case of The Netherlands.

A substantial minority (35%) of the Dutch population is in favor of capital punishment. In this paper, it is argued that in a staunchly abolitionist country such as The Netherlands, the existence and perseverance of such support can be better understood and explained by conceiving of capital punishment support in attitudinal terms as part of a law and order syndrome. Death penalty attitudes are analyzed by means of hierarchic logistic regression analysis. It is shown that support can be modeled quite well, partly in terms of general attitudes to criminal justice, partly in terms of political and sociodemographic parameters. Within the criminal justice attitudes complex, more support is found among those endorsing harsh treatment of offenders, those willing to grant far-reaching powers to justice authorities, those believing that the government is not delivering on the topic of crime fighting, and those who are concerned about the level of crime. Within the political context, more support is enlisted among people who abstain from voting and those who vote at either extreme of the political spectrum as opposed to central parties' supporters. In sociodemographic segments it is the younger and poorly educated who are the strongest supporters of capital punishment. It is suggested that endorsing capital punishment can be better understood as an expressive act, displaying dissatisfaction with judicial and political elites in the country.

Dietary long chain PUFAs differentially affect hippocampal muscarinic 1 and serotonergic 1A receptors in experimental cerebral hypoperfusion.

The chronic dietary intake of essential polyunsaturated fatty acids (PUFAs) can modulate learning and memory by being incorporated into neuronal plasma membranes. Representatives of two PUFA families, the n-3 and n-6 types become integrated into membrane phospholipids, where the actual (n-6)/(n-3) ratio can determine membrane fluidity and thus the function of membrane-bound proteins. In the present experiment we studied hippocampal neurotransmitter receptors after chronic administration of n-3 PUFA enriched diets in a brain hypoperfusion model, which mimics decreased cerebral perfusion as it occurs in ageing and dementia. Male Wistar rats received experimental diets with a decreased (n-6)/(n-3) ratio from weaning on. Chronic experimental cerebral hypoperfusion was imposed by a permanent, bilateral occlusion of the common carotid arteries (2VO) at the age of 4 months. The experiment was terminated when the rats were 7 months old. Three receptor types, the muscarinic 1, serotonergic 1A and the glutaminergic NMDA receptors were labeled in hippocampal slices by autoradiographic methods. Image analysis demonstrated that 2VO increased muscarinic 1 and NMDA receptor density, specifically in the dentate gyrus and the CA3 region, respectively. The increased ratio of n-3 fatty acids in combination with additional dietary supplements enhanced the density of the serotonergic 1A and muscarinic 1 receptors, while n-3 fatty acids alone increased binding only to the muscarinic 1 receptors. Since the examined receptor types reacted differently to the diets, we concluded that besides changes in membrane fluidity, the biochemical regulation of receptor sensitivity might also play a role in increasing hippocampal receptor density.

From moral theory to penal attitudes and back: a theoretically integrated modeling approach.

From a moral standpoint, we would expect the practice of punishment to reflect a solid and commonly shared legitimizing framework. Several moral legal theories explicitly aim to provide such frameworks. Based on the theories of Retributivism, Utilitarianism, and Restorative Justice, this article first sets out to develop a theoretically integrated model of penal attitudes and then explores the extent to which Dutch judges' attitudes to punishment fit the model. Results indicate that penal attitudes can be measured in a meaningful way that is consistent with an integrated approach to moral theory. The general structure of penal attitudes among Dutch judges suggests a streamlined and pragmatic approach to legal punishment that is identifiably founded on the separate concepts central to moral theories of punishment. While Restorative Justice is frequently presented as an alternative paradigm, results show it to be smoothly incorporated within the streamlined approach.