Is That "Mr." or "Ms." Lemon? An Investigation of Grammatical and Semantic Gender on the Perception of Household Odorants.

Linguistic relativism is the idea that the structure of language influences thought. The present study investigates linguistic relativism by asking whether people who speak a gendered language think of objects in a way that is consistent with the grammatical gender (more masculine or feminine) more than people who speak a language without that linguistic feature. Odorants are unique stimuli for examining this question, because they carry a semantic gender without a biological one, yet gender is thought to be a central dimension of their perception. Canadian participants in the present study (native French or native English speakers) described a set of labeled odorants that varied systematically in grammatical and semantic gender to produce an implicit gender measure and also explicitly rated them for gender. Analysis of the implicit femininity contained in participants' descriptions showed a tendency for both native French and native English speakers to produce descriptions that were consistent with semantic gender; moreover, there were no effects of grammatical gender on implicit or explicit ratings. These results suggest that in a bilingual culture, French speakers may have been influenced by the anthropomorphism associated with odorants rather than grammatical gender.

Scent of a Woman-Or Man: Odors Influence Person Knowledge.

First impressions of social traits are regularly, rapidly, and readily determined from limited information about another individual. Relatively little is known about the way that olfactory information, particularly from scents that are not body odors, alters a first impression. Can the attributes of an odorant be conferred onto a person associated with that scent? To explore this, 101 participants were asked to form an impression of a hypothetical person based on the following stimuli: A gender-neutral silhouette, a list of six personal characteristics, and one of five odorants. Participants then rated the likelihood that the hypothetical person possessed each of 51 personality traits that were determined a priori as falling into six attribute categories. Participants also directly rated all odorants for the six categories and intensity. A T-test showed that ratings of the hypothetical person were less disparate from the odor that was presented during impression formation than from other odors. ANOVA revealed that the effects were heterogeneous, with odorants varying in their effectiveness in associating the hypothetical person with categories. The present data suggest that a hypothetical person can be imbued with the specific attributes of an odor and that some odors are better at contributing to impressions than others.

Tracking smell loss to identify healthcare workers with SARS-CoV-2 infection.

INTRODUCTION: Healthcare workers (HCW) treating COVID-19 patients are at high risk for infection and may also spread infection through their contact with vulnerable patients. Smell loss has been associated with SARS-CoV-2 infection, but it is unknown whether monitoring for smell loss can be used to identify asymptomatic infection among high risk individuals. In this study we sought to determine if tracking smell sensitivity and loss using an at-home assessment could identify SARS-CoV-2 infection in HCW. METHODS AND FINDINGS: We performed a prospective cohort study tracking 473 HCW across three months to determine if smell loss could predict SARS-CoV-2 infection in this high-risk group. HCW subjects completed a longitudinal, behavioral at-home assessment of olfaction with household items, as well as detailed symptom surveys that included a parosmia screening questionnaire, and real-time quantitative polymerase chain reaction testing to identify SARS-CoV-2 infection. Our main measures were the prevalence of smell loss in SARS-CoV-2-positive HCW versus SARS-CoV-2-negative HCW, and timing of smell loss relative to SARS-CoV-2 test positivity. SARS-CoV-2 was identified in 17 (3.6%) of 473 HCW. HCW with SARS-CoV-2 infection were more likely to report smell loss than SARS-CoV-2-negative HCW on both the at-home assessment and the screening questionnaire (9/17, 53% vs 105/456, 23%, P < .01). 6/9 (67%) of SARS-CoV-2-positive HCW reporting smell loss reported smell loss prior to having a positive SARS-CoV-2 test, and smell loss was reported a median of two days before testing positive. Neurological symptoms were reported more frequently among SARS-CoV-2-positive HCW who reported smell loss compared to those without smell loss (9/9, 100% vs 3/8, 38%, P < .01). CONCLUSIONS: In this prospective study of HCW, self-reported changes in smell using two different measures were predictive of SARS-CoV-2 infection. Smell loss frequently preceded a positive test and was associated with neurological symptoms.

Tracking Smell Loss to Identify Healthcare Workers with SARS-CoV-2 Infection.

BACKGROUND: Healthcare workers (HCW) treating COVID-19 patients are at high risk for infection and may also spread infection through their contact with vulnerable patients. Smell loss has been associated with SARS-CoV-2 infection, but it is unknown whether monitoring for smell loss can be used to identify asymptomatic infection among high risk individuals, like HCW. METHODS: We performed a prospective cohort study, tracking 473 HCW across three months to determine if smell loss could predict SARS-CoV-2 infection in this high-risk group. HCW subjects completed a longitudinal, novel behavioral at-home assessment of smell function with household items, as well as detailed symptom surveys that included a parosmia screening questionnaire, and RT-qPCR testing to identify SARS-CoV-2 infection. RESULTS: SARS-CoV-2 was identified in 17 (3.6%) of 473 HCW. Among the 17 infected HCW, 53% reported smell loss, and were more likely to report smell loss than COVID-negative HCW on both the at-home assessment and the screening questionnaire (P < .01). 67% reported smell loss prior to having a positive SARS-CoV-2 test, and smell loss was reported a median of two days before testing positive. Neurological symptoms were reported more frequently among COVID-positive HCW who reported smell loss (P < .01). CONCLUSIONS: In this prospective study of HCW, self-reported changes in smell using two different measures were predictive of COVID-19 infection. Smell loss frequently preceded a positive test and was associated with neurological symptoms.

IL-10-Dependent Crosstalk between Murine Marginal Zone B Cells, Macrophages, and CD8α+ Dendritic Cells Promotes Listeria monocytogenes Infection.

Type 1 CD8α+ conventional dendritic cells (cDC1s) are required for CD8+ T cell priming but, paradoxically, promote splenic Listeria monocytogenes infection. Using mice with impaired cDC2 function, we ruled out a role for cDC2s in this process and instead discovered an interleukin-10 (IL-10)-dependent cellular crosstalk in the marginal zone (MZ) that promoted bacterial infection. Mice lacking the guanine nucleotide exchange factor DOCK8 or CD19 lost IL-10-producing MZ B cells and were resistant to Listeria. IL-10 increased intracellular Listeria in cDC1s indirectly by reducing inducible nitric oxide synthase expression early after infection and increasing intracellular Listeria in MZ metallophilic macrophages (MMMs). These MMMs trans-infected cDC1s, which, in turn, transported Listeria into the white pulp to prime CD8+ T cells. However, this also facilitated bacterial expansion. Therefore, IL-10-mediated crosstalk between B cells, macrophages, and cDC1s in the MZ promotes both Listeria infection and CD8+ T cell activation.

Efficacy and Safety of High-Specific-Activity 131I-MIBG Therapy in Patients with Advanced Pheochromocytoma or Paraganglioma.

Patients with metastatic or unresectable (advanced) pheochromocytoma and paraganglioma (PPGL) have poor prognoses and few treatment options. This multicenter, phase 2 trial evaluated the efficacy and safety of high-specific-activity 131I-meta-iodobenzylguanidine (HSA 131I-MIBG) in patients with advanced PPGL. Methods: In this open-label, single-arm study, 81 PPGL patients were screened for enrollment, and 74 received a treatment-planning dose of HSA 131I-MIBG. Of these patients, 68 received at least 1 therapeutic dose (∼18.5 GBq) of HSA 131I-MIBG intravenously. The primary endpoint was the proportion of patients with at least a 50% reduction in baseline antihypertensive medication use lasting at least 6 mo. Secondary endpoints included objective tumor response as assessed by Response Evaluation Criteria in Solid Tumors version 1.0, biochemical tumor marker response, overall survival, and safety. Results: Of the 68 patients who received at least 1 therapeutic dose of HSA 131I-MIBG, 17 (25%; 95% confidence interval, 16%-37%) had a durable reduction in baseline antihypertensive medication use. Among 64 patients with evaluable disease, 59 (92%) had a partial response or stable disease as the best objective response within 12 mo. Decreases in elevated (≥1.5 times the upper limit of normal at baseline) serum chromogranin levels were observed, with confirmed complete and partial responses 12 mo after treatment in 19 of 28 patients (68%). The median overall survival was 36.7 mo (95% confidence interval, 29.9-49.1 mo). The most common treatment-emergent adverse events were nausea, myelosuppression, and fatigue. No patients had drug-related acute hypertensive events during or after the administration of HSA 131I-MIBG. Conclusion: HSA 131I-MIBG offers multiple benefits, including sustained blood pressure control and tumor response in PPGL patients.

Migratory CD11b+ conventional dendritic cells induce T follicular helper cell-dependent antibody responses.

T follicular helper (Tfh) cells are a subset of CD4+ T cells that promote antibody production during vaccination. Conventional dendritic cells (cDCs) efficiently prime Tfh cells; however, conclusions regarding which cDC instructs Tfh cell differentiation have differed between recent studies. We found that these discrepancies might exist because of the unusual sites used for immunization in murine models, which differentially bias which DC subsets access antigen. We used intranasal immunization as a physiologically relevant route of exposure that delivers antigen to all tissue DC subsets. Using a combination of mice in which the function of individual DC subsets is impaired and different antigen formulations, we determined that CD11b+ migratory type 2 cDCs (cDC2s) are necessary and sufficient for Tfh induction. DC-specific deletion of the guanine nucleotide exchange factor DOCK8 resulted in an isolated loss of CD11b+ cDC2, but not CD103+ cDC1, migration to lung-draining lymph nodes. Impaired cDC2 migration or development in DC-specific Dock8 or Irf4 knockout mice, respectively, led to reduced Tfh cell priming, whereas loss of CD103+ cDC1s in Batf3-/- mice did not. Loss of cDC2-dependent Tfh cell priming impaired antibody-mediated protection from live influenza virus challenge. We show that migratory cDC2s uniquely carry antigen into the subanatomic regions of the lymph node where Tfh cell priming occurs-the T-B border. This work identifies the DC subset responsible for Tfh cell-dependent antibody responses, particularly when antigen dose is limiting or is encountered at a mucosal site, which could ultimately inform the formulation and delivery of vaccines.

Non-sclerotic bone involvement in Erdheim-Chester: PET/CT and MRI findings in a 15-year-old boy.

Erdheim-Chester disease is a rare form of non-Langerhans cell histiocytosis with multi-organ infiltration that occurs mainly in adults. Pediatric cases are extremely rare. Here we report a case of multisystemic Erdheim-Chester disease in a 15-year-old boy with central nervous system involvement and skeletal findings. Positron emission tomography (PET) and MRI were used to demonstrate characteristic bilateral, symmetrical medullary involvement of the metadiaphyses of long bones in the absence of the classic sclerotic radiographic appearance. This illustrates the potential for earlier diagnosis and visualization of therapeutic response in children.

Metacognitive knowledge of olfactory dysfunction in Parkinson's disease.

It is well known that patients with Parkinson's Disease (PD) suffer from olfactory impairments, but it is not clear whether patients are aware of their level of deficit in olfactory functioning. Since PD is a neurodegenerative disorder and its progression may be correlated with olfactory loss (Ansari & Johnson, 1975; but see also Doty, Deems, & Stellar, 1988), it is possible that these patients would be subject to metacognitive errors of over-estimation of olfactory ability (White & Kurtz, 2003). Nineteen non-demented PD patients and 19 age-matched controls were each given an objective measure of olfactory identification (the UPSIT, Doty, Shaman, Kimmelman, & Dann, 1984) and a subjective measure involving a questionnaire that asked them to self-rate both their olfactory function generally and their ability to smell each of 20 odors, 12 of which were assessed on the UPSIT. All of the PD patients showed impaired olfactory ability, as did 7 of the controls, according to the UPSIT norms. Self-rated and performance-based olfactory ability scores were significantly correlated in controls (r=.49, p=.03) but not in patients with PD (r=.20, p=.39). When the 12 odors common to both the self-rated questionnaire and UPSIT were compared, PD patients were less accurate than controls (t(36)=-4.96, p<.01) at estimating their own ability and the number of over-estimation errors was significantly higher (tone-tailed(29)=1.80, p=.04) in PD patients than in the control group, showing less metacognitive awareness of their ability than controls. These results support the idea that olfactory metacognition is often impaired in PD, as well as in controls recruited for normosmic ability (Wehling, Nordin, Espeseth, Reinvang, & Lundervold, 2011), and indicate that people with PD generally exhibit over-estimation of their olfactory ability at a rate that is higher than controls. These findings imply that PD patients, unaware of their olfactory deficit, are at greater risk of harm normally detected through olfaction, such as smoke or spoiled foods.

Mutation of the Xylanase regulator 1 causes a glucose blind hydrolase expressing phenotype in industrially used Trichoderma strains.

BACKGROUND: Trichoderma reesei is an organism involved in degradation of (hemi)cellulosic biomass. Consequently, the corresponding enzymes are commonly used in different types of industries, and recently gained considerable importance for production of second-generation biofuel. Many industrial T. reesei strains currently in use are derived from strain Rut-C30, in which cellulase and hemicellulase expression is released from carbon catabolite repression. Nevertheless, inducing substances are still necessary for a satisfactory amount of protein formation. RESULTS: Here, we report on a T. reesei strain, which exhibits a very high level of xylanase expression regardless if inducing substances (e.g. D-xylose, xylobiose) are used. We found that a single point mutation in the gene encoding the Xylanase regulator 1 (Xyr1) is responsible for this strong deregulation of endo-xylanase expression and, moreover, a highly elevated basal level of cellulase expression. This point mutation is localized in a domain that is common in binuclear zinc cluster transcription factors. Only the use of sophorose as inducer still leads to a slight induction of cellulase expression. Under all tested conditions, the formation of cbh1 and cbh2 transcript level strictly follows the transcript levels of xyr1. The correlation of xyr1 transcript levels and cbh1/cbh2 transcript levels and also their inducibility via sophorose is not restricted to this strain, but occurs in all ancestor strains up to the wild-type QM6a. CONCLUSIONS: Engineering a key transcription factor of a target regulon seems to be a promising strategy in order to increase enzymes yields independent of the used substrate or inducer. The regulatory domain where the described mutation is located is certainly an interesting research target for all organisms that also depend so far on certain inducing conditions.

Exploring the Sexuality of African American Older Women.

AIMS: To identify sexually-related themes of the sexuality of older African American women. STUDY DESIGN: Mixed method. PLACE AND DURATION OF STUDY: Department of Psychology, California State University Northridge, between July 2009 and June 2011. METHODOLOGY: We included 13 African American older women (57 to 82 years of age), 11 of whom self-identified as heterosexual, one as bisexual, and one as lesbian. We used a semi-structured interview protocol through which we explored some aspects of the respondents' sexuality (assessed at a superficial level, to be as tactful as possible). Moreover, we collected information on demographics and self-rated physical health. Two co-authors served as coders, and used content analysis to identify the most salient sexuality themes. RESULTS: Emerging themes were (in order from most to least endorsed): having sexual desire (often unfulfilled); engaging in less sexual activity in older age; experiencing changes in one's sexual life as a function of absence of a spouse; and exercising control over how one's sexual life is conducted. Motivated by the paucity of our sexuality data, we have also provided suggestions to scholars interested in conducting more in-depth further research on this topic with older African American women. CONCLUSION: Our findings indicate that the common notion that older women are asexual is a myth, while lack of a suitable sexual partner is a problem reported by many African American older women who would otherwise enjoy sexual interaction.

Adding cocoa to sucrose: the effect on cold pain tolerance.

The sweet taste of sucrose acts as an analgesic, whereas the taste of a bitter substance decreases pain tolerance. The present experiment explores the analgesic effect of a complex taste and asks how adding cocoa, a substance often associated with sweet foods but that has a bitter taste, to a sucrose solution affects cold pain tolerance. The 24 male participants were exposed to Cold Pressor Tests (CPTs) while holding 1 of 3 tastants in their mouths: water, sucrose, or sucrose with cocoa added. After each CPT, participants rated pain intensity and tastant qualities. Intraoral sucrose increased the amount of time that men were able to leave their hands in cold water, whereas the cocoa solution did not. Solutions did not differ in pleasantness or sweetness, but the cocoa solution was rated as more bitter. Bitterness ratings of cocoa exceeded the ratings of sucrose (corrected for water) by an average of 16.9% (P = 0.02), which, in turn, produced a 30% reduction in the duration of pain tolerance (P = 0.002). These results suggest that the addition of a bitter substance reduces cues to the nutritive value of sucrose that may drive its analgesic effect.

Identification of a gene coding for a deglycosylating enzyme in Hypocrea jecorina.

An enzyme with mannosyl glycoprotein endo-N-acetyl-beta-D-glucosaminidase (ENGase)-type activity was partially purified from the extracellular medium of the mould Hypocrea jecorina (Trichoderma reesei). Internal peptides were generated and used to identify the gene in the T. reesei genome. The active enzyme is processed both at the N- and at the C-terminus. High-mannose-type glycoproteins are good substrates, whereas complex-type glycans are not hydrolysed. The enzyme represents the first fungal member of glycoside hydrolase family 18 with ENGase-type activity. Bacterial ENGases and the fungal chitinases belonging to the same family show very low homology with Endo T. Database searches identify several highly homologous genes in fungi and the activity is also found within other Trichoderma species. This ENGase activity, not coregulated with cellulase production, could be responsible for the extensive N-deglycosylation observed for several T. reesei cellulases.

Design of a novel Couette flow bioreactor to study the growth of fungal microorganism.

Cultivations using Trichoderma reesei Rut C-30 were performed in a 5-l Couette flow bioreactor (CFB) which was designed and built to perform experiments in batch and continuous modes. Process parameters such as dissolved oxygen, pH and temperature were measured and controlled without disturbing the shear profile inside the bioreactor. Effect of shear on the growth, protein production and morphology was studied by performing runs at 100, 200, 300 and 400 rpm. At higher shear rates, lower protein production rate and activity, and higher rate of fragmentation were observed. Also, the cell thickness decreased with increasing speed, going from 8.3 microm for the experiment at 100 rpm to 4.3 microm at 400 rpm. The effect of substrate, lactose (an inducer) or glucose, was investigated by switching the feed medium during the two runs performed at 300 and 400 rpm. The novel design of the CFB used in the present study includes a large volume that allows growing larger size microorganisms (e.g. fungi) and permits larger sampling volumes without affecting the cultivation. It also has the ability to carry out experiments for long periods of time, both in batch and continuous modes.

A second look at the structure of human olfactory memory.

How do we remember olfactory information? Is the architecture of human olfactory memory unique compared with that of memory for other types of stimuli? Ten years ago, a review article evaluated these questions, as well as the distinction between long- and short-term olfactory memory, with three lines of evidence: capacity differences, coding differences, and neuropsychological evidence, though serial position effects were also considered. From the data available at the time, the article preliminarily suggested that olfactory memory was a two-component system that was not qualitatively different from memory systems for other types of stimuli. The decade that has elapsed since then has ushered in considerable changes in theories of memory structure and provided huge advances in neuroscience capabilities. Not only have many studies exploring various aspects of olfactory memory been published, but a model of olfactory perception that includes an integral unitary memory system also has been presented. Consequently, the structure of olfactory memory is reevaluated in the light of further information currently available with the same theoretical lines of evidence previously considered. This evaluation finds that the preponderance of evidence suggests that, as in memory for other types of sensory stimuli, the short-term-long-term distinction remains a valuable dissociation for conceptualizing olfactory memory, though perhaps not as architecturally separate systems.

The relationship between PROP and ethanol preferences: an evaluation of 4 inbred mouse strains.

Ethanol's taste attributes undoubtedly contribute to the development of drug preference. Ethanol's taste is both sweet and bitter. Taster status for bitter 6-n-propylthiouracil (PROP) has been proposed as a genetic marker for alcoholism; however, human results are conflicting. We collected preference scores for both tastants in 4 mouse strains selected on the basis of previously reported taste preference, with the generally accepted idea that inbred mice show minimal within-strain variation. Eighty-eight male mice (22 per strain) participated. The strains were as follows: C57BL/6J, ethanol preferring; BALB/cJ, ethanol avoiding; SWR/J, PROP avoiding; and C3HeB/FeJ, PROP neutral. Using a brief-access (1-min trials) 2-bottle preference test, we assessed the taste response of each strain to PROP and ethanol on separate days. Although PROP avoiding versus neutral mice could be segregated into significantly different populations, this was not the case for ethanol avoiding versus preferring mice, and all strains showed high variability. On average, only BALB/cJ, SWR/J, and C3HeB/FeJ mice conformed to their literature-reported preferences; nonetheless, there were a substantial number of discordant animals. C57BL/6J did not conform to previous results, indicating that they are ethanol preferring. Finally, we did not observe a significant relationship between PROP and ethanol preferences across strains. The high variability per strain and the number of animals in disagreement with their respective literature-reported preference raise concerns regarding their utility for investigations underlying mechanisms of taste-mediated ingestive responses. Absent postingestive consequences, the brief-access results suggest a possible degree of previously masked polymorphisms in taste preferences or a more recent drift in underlying genetic factors. The absence of a relationship between PROP and ethanol indicates that the bitter quality in ethanol may be more highly related to other bitter compounds that are mediated by different genetic influences.

Chemosensory cross-modal stroop effects: congruent odors facilitate taste identification.

In order to explore the cross-modal cognitive associations between smell and taste, a chemosensory analogue of the Stroop task (Stroop 1935) was developed. Fourteen participants were presented with an odorant and a tastant and asked to identify the tastant as "sweet" or "sour" by pressing 1 of 2 buttons as quickly as possible. Participants were faster to name the taste when it was presented with an odor that was congruent (e.g., strawberry/sweet) than with an incongruent odor (e.g., strawberry/sour). These results support the concept of a high level of cognitive integration between the senses of smell and taste and illustrates occasions of interference between information arising from different sensory systems.

The effect of NMDA-NR2B receptor subunit over-expression on olfactory memory task performance in the mouse.

The N-methyl-D-aspartate (NMDA) receptor in the forebrain is thought to modulate some forms of memory formation, with the NR2B subunit being particularly relevant to this process. Relative to wild-type mice, transgenic animals in which the NR2B subunit was over-expressed demonstrate superior memory in a number of behavioral tasks, including object recognition [Nature 401 (1999) 63]. The purpose of the present study was to explore the generality of such phenomena, interpreted as the effect of increasing NR2B expression on the retention of other types of sensory-related information. To accomplish this, we focused our evaluation on the highly salient sensory modality of olfaction. In the first experiment, mice performed both a novel-object-recognition task identical to that performed by Tang et al. [Nature 401 (1999) 63] and a novel-odor-recognition task analogously constructed. Although the results of the object recognition task were consistent with the previous literature, there was no evidence of an effect of NR2B over-expression on the retention of odor recognition memory in the specific task performed. As it was possible that, unlike object recognition memory, novel odor recognition is not NMDA-receptor-dependent, a second task was designed using the social transmission of food preference paradigm. In contrast to the foregoing olfactory task, there is evidence that the latter procedure is, indeed, NMDA-dependent. The results of the second study demonstrated that transgenic mice with NR2B over-expression had a clear memory advantage in this alternative odor memory paradigm. Taken together, these results suggest the NR2B subunit is an important component in some but not all forms of olfactory memory organization. Moreover, for those functions that are NMDA-receptor-dependent, these data support the growing literature demonstrating the importance of the NR2B subunit.

Effectiveness of gabapentin for treatment of burning mouth syndrome.

Burning mouth syndrome is a debilitating disorder involving oral pain that may have at least 4 underlying causes. Although several treatments have been proposed, none seems to be universally effective. We report the case of a 67-year-old woman with unremitting oral burning that is increased with the application of anesthetic agents. Initial treatments with nortriptyline hydrochloride and sertraline hydrochloride were contraindicated because of adverse effects, but the administration of gabapentin significantly reduced oral burning. The present case illustrates the effectiveness of gabapentin as a treatment of burning mouth syndrome.

The relationship between metacognitive awareness of olfactory ability and age in people reporting chemosensory disturbances.

When determining how much to rely on olfactory information, accurate estimates of olfactory ability are important. Nevertheless, metacognitive errors regarding olfactory ability are common in the normal population. Are older adults less accurate at estimating olfactory ability, or do they simply tend to make a particular type of estimation error? We examined data from 203 patients, all of whom self-rated olfactory ability and then were tested with the Odorant Confusion Matrix. The patients' self-reports did not match the objective measure of olfactory ability in 42% of the cases, thus demonstrating a high level of impairment in metacognitive knowledge. Neither the cause of loss nor the patient's age was related to the inability to accurately estimate olfactory loss, but both contributed to the type of metacognitive errors patients made. Younger patients tended to underestimate their olfactory ability, thus behaving similarly to patients with "blindsight" who demonstrate residual visual function in the absence of awareness. Older patients tended to overestimate their olfactory ability, thus making the metacognitive error of deficit unawareness that is observed in a number of other clinical syndromes. These findings indicate that older patients are not necessarily more likely to make errors in estimating their olfactory ability; rather, they are more prone to a particular type of metacognitive error than are younger patients.