Assessment of Hydration Status Using Conventional Method and Salivary Osmolarity as a Point-of-care Tool.

Dehydration is a well-known problem worldwide, and its assessment can be challenging due to confusing physical signs. The most effective way to assess hydration status is through the costly stable isotope methodology, but this approach has practical limitations. More commonly accepted and utilized indicators of hydration status are hematological and urinary parameters. However, hematological markers require invasive methods, and urinary markers have varying degrees of success in tracking hydration changes. While alterations in body weight can serve as a means of promptly evaluating hydration status, various factors such as food consumption, fluid intake, fecal losses, and urine production can impact these changes. Researchers have turned their attention to saliva as a potential marker and point-of-care (POC) testing to address the limitations of existing biomarkers. Saliva is appealing due to its easy collection process and similarities to extracellular fluid in terms of water and ion concentrations. Recent studies have shown that saliva flow rate, osmolarity/osmolality, and total protein concentration can effectively monitor changes in body mass during acute dehydration. Misdiagnosing dehydration can have severe clinical consequences, leading to morbidity and even mortality. This narrative review focuses on recognizing the significance of hydration assessment, monitoring, and the potential of salivary osmolarity (SOSM) as an assessment tool. Healthcare professionals can improve their practices and interventions to optimize hydration and promote overall wellness using such tools.

Differences in urine creatinine and osmolality between black and white Americans after accounting for age, moisture intake, urine volume, and socioeconomic status.

Urine osmolality is used throughout research to determine hydration levels. Prior studies have found black individuals to have elevated urine creatinine and osmolality, but it remains unclear which factors explain these findings. This cross-sectional, observational study sought to understand the relationship of self-reported race to urine creatinine and urine osmolality after accounting for age, socioeconomic status, and fluid intake. Data from 1,386 participants of the 2009-2012 National Health and Nutrition Examination Survey were utilized. Age, poverty-to-income ratio (PIR), urine flow rate (UFR), fluid intake, estimated lean body mass (LBM), urine creatinine, and urine osmolality were measured. In a sex-specific manner, black and white participants were matched on age, dietary moisture, UFR, and PIR. Urine creatinine was greater in black men (171 mg/dL) than white men (150 mg/dL) and greater in black women (147 mg/dL) than white women (108 mg/dL) (p < .001). Similarly, urine osmolality was greater in black women than white women (723 vs. 656 mOsm/kg, p = .001), but no difference was observed between white and black men (737 vs. 731 mOsm/kg, p = .417). Estimated LBM was greater in black men (61.8 kg) and women (45.5 kg) than in white men (58.9 kg) and women (42.2 kg) (p≤.001). The strongest correlate of urine osmolality in all race-sex groups was urine creatinine (Spearman ρ = .68-.75). These results affirm that individuals identifying as black produce higher urine creatinine concentrations and, in women, higher urine osmolality after matching for age, fluid intake, and socioeconomic status. The findings suggest caution when comparing urine hydration markers between racial groups.

End-of-day assessment of asymptomatic versus highly symptomatic soft contact lens wearers.

PURPOSE: To investigate differences in key clinical parameters between asymptomatic and highly symptomatic soft contact lens (CL) wearers after 14 h of wear. METHODS: In this pilot investigation, Phase 1 identified asymptomatic (CLDEQ-8 score ≤ 7) and highly symptomatic (CLDEQ-8 score ≥ 20) subjects after fitting with nelfilcon A CLs. Phase 2 investigated the following over a single nelfilcon A CL-wearing day (14 ± 2 h): blinking characteristics, tear meniscus height (TMH), non-invasive tear break-up time (NIBUT), tear film osmolarity and eyelid margin staining. Parameters for the two groups were compared using linear mixed models and post-hoc testing. The relationship between comfort scores and the clinical parameters was also investigated. RESULTS: Overall, 161 and 42 subjects were enrolled into Phase 1 and 2, respectively. Twenty-five asymptomatic and 17 symptomatic subjects completed Phase 2. Lower eyelid TMH was decreased after 14 h in symptomatic compared with asymptomatic subjects (least square mean [LSM] difference -0.04 mm, 95% CI: -0.07, -0.01). Osmolarity was lower in symptomatic than in asymptomatic subjects at fitting (LSM difference -9.89, 95% CI: -18.91, -0.86). Upper eyelid margin staining was greater after 14 h in symptomatic than in asymptomatic subjects (LSM difference 0.53, 95% CI: 0.01, 1.05) and greater after 14 h than baseline in the symptomatic group (LSM difference 0.61, 95% CI: 0.16, 1.07). There was a significant relationship between comfort and upper eyelid margin staining (r = -0.40, 95% CI: -0.63, -0.11) and blink rate (r = -0.31, 95% CI: -0.57, -0.003). CONCLUSION: The potential parameters most effective in differentiating asymptomatic from symptomatic wearers were upper eyelid margin staining and lower TMH. The parameter with the strongest relationship to comfort was upper eyelid margin staining, where higher comfort scores were associated with lower levels of staining.

The weight, urine colour and thirst Venn diagram is an accurate tool compared with urinary and blood markers for hydration assessment at morning and afternoon timepoints in euhydrated and free-living individuals.

The weight, urine colour and thirst (WUT) Venn diagram is a practical hydration assessment tool; however, it has only been investigated during first-morning. This study investigated accuracy of the WUT Venn diagram at morning and afternoon timepoints compared with blood and urine markers. Twelve men (21 ± 2 years; 81·0 ± 15·9 kg) and twelve women (22 ± 3 years; 68·8 ± 15·2 kg) completed the study. Body mass, urine colour, urine specific gravity (USG), urine osmolality (UOSM), thirst and plasma osmolality (POSM) were collected at first-morning and afternoon for 3 consecutive days in free-living (FL) and euhydrated states. Number of markers indicating dehydration levels were categorised into either 3, 2, 1 or 0 WUT markers. Receiver operating characteristics analysis calculated the sensitivity and specificity of 1, 2 or 3 hydration markers in detecting dehydration or euhydration. Specificity values across morning and afternoon exhibited high diagnostic accuracy for USG (0·890-1·000), UOSM (0·869-1·000) and POSM (0·787-0·990) when 2 and 3 WUT markers were met. Sensitivity values across both timepoints exhibited high diagnostic accuracy for USG (0·826-0·941) and UOSM (0·826-0·941), but not POSM in the afternoon (0·324) when 0 and 1 WUT markers were met. The WUT Venn diagram is accurate in detecting dehydration for WUT2 and WUT3 based off USG, UOSM and POSM during first-morning and afternoon. Applied medical, sport and occupational practitioners can use this tool in field settings for hydration assessment not only at various timepoints throughout the day but also in FL individuals.

Cross-sectional analysis of racial differences in hydration and neighborhood deprivation in young adults.

BACKGROUND: Inadequate hydration is associated with cardiovascular and kidney disease morbidity and all-cause mortality. Compared with White individuals, Black individuals exhibit a higher prevalence of inadequate hydration, which may contribute to racial health disparities. However, the underlying reasons for these differences in hydration remain unclear. OBJECTIVE: This cross-sectional study aimed to investigate whether neighborhood deprivation contributes to racial differences in hydration status. METHODS: We assessed 24 Black and 30 White college students, measuring 24-hour urine osmolality, urine flow rate, urine specific gravity, and plasma copeptin concentration. Participants recorded their food and fluid intake for 3 d to assess total water intake from food and beverages. Neighborhood socioeconomic deprivation was measured using a tract-level Area Deprivation Index. RESULTS: Black participants exhibited higher urine osmolality (640 [314] compared with 440 [283] mOsm/kg H2O, respectively, P = 0.006) and lower urine flow rate (1.06 [0.65] compared with 1.71 [0.89] ml/min, respectively, P = 0.009) compared with White participants, indicating greater hypohydration among Black participants. Black participants reported lower total water intake from food and beverages than White participants (2.3 ± 0.7 compared with 3.5 ± 1.1 L/day, respectively, P < 0.01). Black participants exhibited higher copeptin than White participants (6.3 [3.1] compared with 4.5 [2.3] pmol/L, P = 0.046), and urine osmolality mediated 67% of the difference (P = 0.027). Black participants reported greater cumulative exposure to neighborhood deprivation during childhood (ages 0-18 y). Furthermore, neighborhood deprivation during childhood was associated with urine specific gravity (P = 0.031) and total water intake from food and beverages (P = 0.042) but did not mediate the racial differences in these measures. CONCLUSION: Our data suggest that compared with White young adults, Black young adults are hypohydrated and exhibit higher plasma copeptin concentration, and that greater neighborhood deprivation is associated with chronic underhydration irrespective of race. This trial was registered at clinicaltrials.gov as NCT04576338.

Hydration Status Assessment in Older Patients.

BACKGROUND: Hydration disturbances are common in old age: the reported prevalence of dehydration in elderly patients ranges from 19% to 89%, depending on the definition and the population in question. However, the clinical assessment of patients' hydration status is difficult. In this review, we discuss the diagnostic value of currently used methods that may or may not be suitable for assessing older patients' hydration status. METHODS: We conducted a selective literature search for relevant studies concerning patients aged 65 and above. Of the 355 articles retrieved by the initial search, a multistep selection process yielded 30 that were suitable for inclusion in this review. RESULTS: 107 different methods for the diagnostic assessment of dehydration in older persons were evaluated on the basis of the reviewed publications. High diagnostic value, especially for the determination of hyperosmolar dehydration, was found for serum osmolality, serum sodium concentration, inferior vena cava ultrasonography, a history (from the patient or another informant) of not drinking between meals, and axillary dryness. On the other hand, a variety of clinical signs such as a positive skin turgor test, sunken eyes, dry mouth, tachycardia, orthostatic dysregulation, and dark urine were found to be of inadequate diagnostic value. CONCLUSION: Only five of the 107 methods considered appear to be suitable for determining that a patient is dehydrated. Thus, the available scientific evidence indicates that all clinicians should critically reconsider their own techniques for assessing hydration status in elderly patients. To optimize the clinical assessment of patients' hydration status, there seems to be a need for the rejection of unsuitable methods in favor of either newly developed criteria or of a combination of the best criteria already in use.

Retrospective rationalization of disparities between the concentration dependence of diffusion coefficients obtained by boundary spreading and dynamic light scattering.

This study establishes the existence of substantial agreement between published results from traditional boundary spreading measurements (including synthetic boundary measurements in the analytical ultracenrifuge) on two globular proteins (bovine serum albumin, ovalbumin) and the concentration dependence of diffusion coefficient predicted for experiments conducted under the operative thermodynamic constraints of constant temperature and solvent chemical potential. Although slight negative concentration dependence of the translational diffusion coefficient is the experimentally observed as well as theoretically predicted, the extent of the concentration dependence is within the limits of experimental uncertainty inherent in diffusion coefficient measurement. Attention is then directed toward the ionic strength dependence of the concentration dependence coefficient ([Formula: see text]) describing diffusion coefficients obtained by dynamic light scattering, where, in principle, the operative thermodynamic constraints of constant temperature and pressure preclude consideration of results in terms of single-solute theory. Nevertheless, good agreement between predicted and published experimental ionic strength dependencies of [Formula: see text] for lysozyme and an immunoglobulin is observed by a minor adaptation of the theoretical treatment to accommodate the fact that thermodynamic activity is monitored on the molal concentration scale because of the constraint of constant pressure that pertains in dynamic light scattering experiments.

Association of Tear Osmolarity With Signs and Symptoms of Dry Eye Disease in the Dry Eye Assessment and Management (DREAM) Study.

Purpose: To determine the relationships of (1) tear osmolarity (TO) levels with the severity of signs and symptoms of dry eye disease (DED) and (2) changes in TO with changes in signs and symptoms. Methods: Patients (N = 405) with moderate to severe DED in the Dry Eye Assessment and Management (DREAM) Study were evaluated at baseline and at six and 12 months. Associations of TO with signs and symptoms were evaluated using Pearson correlation coefficient (r) and regression models. Results: The mean (standard deviation [SD]) TO was 303 (16) mOsm/L at baseline and 303 (18) mOsm/L at both six and 12 months. TO was higher in older patients (306 mOsm/L for ≥70 years vs. 300 mOsm/L for <50 years; P = 0.01) and those with Sjögren's disease (311 vs. 302 mOsm/L; P < 0.0001). TO did not differ between patients randomized to placebo and omega-3 fatty acid supplementation. TO was weakly correlated with conjunctival (r = 0.18; P < 0.001) and corneal staining scores (r = 0.17; P < 0.001), tear film break-up time (r = 0.06; P = 0.03), and Schirmer test score (r = -0.07; P = 0.02) but not with Ocular Surface Disease Index scores (r = 0.03; P = 0.40). Changes in signs and were not significantly correlated with change in TO at six or 12 months. Conclusions: Within DREAM, TO was weakly correlated with DED signs, explaining <5% variability in signs. Changes in tear osmolarity were not associated with changes in signs and symptoms of DED, indicating that the association may not be causal.

Prevalence of tear film hyperosmolarity in 1150 patients presenting for refractive surgery assessment.

PURPOSE: To present an analysis of tear film hyperosmolarity in a large, consecutive population and evaluate the correlation of ocular and systemic conditions with tear film osmolarity (TFO). SETTING: Private practice, Sydney, Australia. DESIGN: Single-center, retrospective, consecutive cohort. METHOD: Patients undergoing screening for laser refractive surgery from October 2017 to October 2020 were retrospectively reviewed. 1404 patients (n = 1357 standard, n = 47 postrefractive) undergoing screening for laser refractive surgery from October 2017 to October 2020 were reviewed. Routine examination included TFO and Ocular Surface Disease Index (OSDI) questionnaire. TFO was conducted prior to further tests, and patients refrained from topical eyedrops minimum 2 hours before the appointment. RESULTS: 1404 patients (n = 1357 standards, n = 47 postrefractive) patients were reviewed. Mean highest TFO in the standard population was 299.12 ± 11.94 mOsm/L, with 82.3% of eyes <308 mOsm/L indicating normal tear film homeostasis. The mean intereye TFO difference was 8.17 ± 8.60 mOsm/L, with 65.2% of eyes ≤8 mOsm/L. Mean highest TFO in the postrefractive subgroup was 299.72 ± 11.00 mOsm/L, with a mean intereye difference of 9.02 ± 6.92 mOsm/L. Postrefractive surgery patients indicated higher mean OSDI values of 15.28 ± 14.46 compared with the remainder of the population 9.69 ± 10.56 (P = .012). Significant correlation was demonstrated between TFO scores and OSDI normal classification in the standard population only (P = .005, r = 0.077). The use of contact lens correlated inversely with TFO and OSDI scores (P = .000, r = -0.136, and P = .000, r = -0.152, respectively). CONCLUSIONS: To the authors' knowledge, this study represents the largest available cohort of TFO scores in a standard population presenting for refractive surgery. Although most patients were found to fall within normal ranges, a reasonable percentage were diagnosed with tear hyperosmolarity and therefore at risk for dry eye disease.

Enhancing the population of the encounter complex affects protein complex formation efficiency.

Optimal charge distribution is considered to be important for efficient formation of protein complexes. Electrostatic interactions guide encounter complex formation that precedes the formation of an active protein complex. However, disturbing the optimized distribution by introduction of extra charged patches on cytochrome c peroxidase does not lead to a reduction in productive encounters with its partner cytochrome c. To test whether a complex with a high population of encounter complex is more easily affected by suboptimal charge distribution, the interactions of cytochrome c mutant R13A with wild-type cytochrome c peroxidase and a variant with an additional negative patch were studied. The complex of the peroxidase and cytochrome c R13A was reported to have an encounter state population of 80%, compared to 30% for the wild-type cytochrome c. NMR analysis confirms the dynamic nature of the interaction and demonstrates that the mutant cytochrome c samples the introduced negative patch. Kinetic experiments show that productive complex formation is fivefold to sevenfold slower at moderate and high ionic strength values for cytochrome c R13A but the association rate is not affected by the additional negative patch on cytochrome c peroxidase, showing that the total charge on the protein surface can compensate for less optimal charge distribution. At low ionic strength (44 mm), the association with the mutant cytochrome c reaches the same high rates as found for wild-type cytochrome c, approaching the diffusion limit.

Assessment of tear film osmolarity using the IPen® Vet osmometer in Pug and Shih-Tzu dogs with and without keratoconjunctivitis sicca.

OBJECTIVE: To establish tear film osmolarity (TFO) values in Pugs and Shih-Tzus, with and without keratoconjunctivitis sicca (KCS). ANIMALS STUDIED: A total of 82 adult dogs were evaluated. PROCEDURE: The inclusion criteria for the healthy group was a Schirmer tear test (STT-1) ≥15 mm/min with no clinical signs of KCS, whereas those with KCS had clinical signs and a STT-1 ≤10 mm/min. All animals underwent complete ophthalmological evaluation prior to STT-1 and TFO. Student's t tests were used to compare STT-1 and TFO in KCS and healthy eyes as well as possible differences in TFO between breeds. In addition, a linear regression to model the relationship between the two variables (STT-1 and TFO) was performed. A P-value ≤ 0.05 was considered statistically significant. RESULTS: STT-1 results were significantly lower (p = 0.0001) in the KCS group (4.46 ± 1.74) compared with the control group (18.80 ± 2.02). Mean TFO results were significantly higher in the KCS group (353.02 ± 16.58 mOsm/L) (p < 0.0001) compared with the control group (315.27 ± 6.15 mOsm/L). The formula Y = 365.059-2.625 * X significantly predicts (p < 0.001) the value of the variable Y (TFO mOsm/L) as a function of the variable X (STT-1 mm/min), with a coefficient of determination of 0.71. CONCLUSIONS: The results revealed differences in TFO and STT-1 between KCS and healthy dogs. Additionally, STT-1 and TFO values were correlated with the aim to use STT-1 values to predict TFO values in brachycephalic breeds.

Osmoregulatory power influences tissue ionic composition after salinity acclimation in aquatic decapods.

Decapod crustaceans show variable degrees of euryhalinity and osmoregulatory capacity, by responding to salinity changes through anisosmotic extracellular regulation and/or cell volume regulation. Cell volume regulatory mechanisms involve exchange of inorganic ions between extra- and intra-cellular (tissue) compartments. Here, this interplay of inorganic ions between both compartments has been evaluated in four decapod species with distinct habitats and osmoregulatory strategies. The marine/estuarine species Litopenaeus vannamei (Lv) and Callinectes danae (Cd) were submitted to reduced salinity (15‰), after acclimation to 25 and 30‰, respectively. The freshwater Macrobrachium acanthurus (Ma) and Aegla schmitti (As) were submitted to increased salinity (25‰). The four species were salinity-challenged for both 5 and 10 days. Hemolymph osmolality, sodium, chloride, potassium, and magnesium were assayed. The same inorganic ions were quantified in muscle samples. Muscle hydration (MH) and ninhydrin-positive substances (NPS) were also determined. Lv showed slight hemolymph dilution, increased MH and no osmotically-relevant decreases in muscle osmolytes; Cd displayed hemolymph dilution, decreased muscular NaCl and stable MH; Ma showed hypo-regulation and steady MH, with no change in muscle ions; As conformed hemolymph sodium but hypo-regulated chloride, had stable MH and increased muscle NPS and ion levels. Hemolymph and muscle ions (especially chloride) of As were highly correlated (Pearson, +0.83). Significant exchanges between hemolymph and muscle ionic pools were more evident in the two species with comparatively less AER regulatory power, C. danae and A. schmitti. Our findings endorse that the interplay between extracellular and tissue ionic pools is especially detectable in euryhaline species with relatively lower osmoregulatory strength.

Athletes' Self-Assessment of Urine Color Using Two Color Charts to Determine Urine Concentration.

Our objective was to determine self-reported accuracy of an athletic population using two different urine color (Uc) charts (8-color vs. 7-color Uc chart). After approval by the Institutional Review Board, members of an athletic population (n = 189, 20 (19-22) year old student- or tactical athletes and coaches, with n = 99 males and n = 90 females) scored their Uc using two charts. To determine the diagnostic value of Uc, results were compared with urine concentration (osmolality and urine specific gravity, USG). Uc was scored slightly darker with the 8-color vs. 7-color Uc chart (2.2 ± 1.2 vs. 2.0 ± 1.2, respectively, p < 0.001), with a moderate correlation between charts (r = 0.76, 95% CI: 0.69-0.81). Bland-Altman analysis showed a weak reporting bias (r = 0.15, p = 0.04). The area under the curve for correct urine sample classification ranged between 0.74 and 0.86. Higher accuracy for both methods was found when Uc scores were compared to USG over osmolality, indicated by 4.8-14.8% range in difference between methods. The optimal Uc cut-off value to assess a low vs. a high urine concentration for both Uc charts varied in this study between 1 and ≤2 while accuracy for charts was similar up to 77% when compared to USG.

The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method.

Flow cytometry (FC) analysis of erythrocyte shape and related biomechanical properties, such as osmotic fragility, have not moved from a research tool to regular clinical testing. The main reason is existing evidence that various pre-analytical factors influence the mathematical interpretation of the data obtained. With an aim to contribute to the standardization and broaden the use of FC for human erythrocyte shape assessment, freshly prepared peripheral blood erythrocytes isolated from healthy donors were incubated in iso and hypo-osmotic solutions (pure saline, saline with potassium and calcium, and phosphate buffered saline) and examined by FC using values of forward scatter (FSC) and side scatter (SSC). Kurtosis, skewness, Pearson's second skewness coefficient of dissymmetry (PCD), and spherical index, calculated from FSC distributions, were used for the erythrocyte shape evaluation. In all isotonic media FSC distribution and FSC-based morphology parameters showed huge inter-individual and inter-medium variation. With decreasing osmolality, in all media and samples, the size of the erythrocytes increased, and swelling index and kurtosis decreased. However, changes in skewness and PCD were influenced by the medium used and the sample tested. Compared to FSC, SSC signal in isotonic and its change in hypotonic media showed lower inter-individual variation and was not influenced by the type of medium. We propose a spherical index and kurtosis as FSC-based indicators of erythrocyte shape. As more resistant to the influence of the preanalytical treatment, SSC data appeared to be unfairly neglected for the assessment of erythrocyte shape, in comparison to the usually employed FSC data.

Systemic Conditions Associated with Severity of Dry Eye Signs and Symptoms in the Dry Eye Assessment and Management Study.

PURPOSE: Certain systemic conditions are reported to be risk factors for dry eye disease (DED), but their associations with DED severity are not well studied. We evaluated whether systemic conditions reported to be DED risk factors are associated with severity of DED signs and symptoms. DESIGN: Secondary analysis of data from the Dry Eye Assessment and Management Study, a large-scale multicenter randomized clinical trial of patients with moderate to severe DED. PARTICIPANTS: Five hundred thirty-five adult patients with moderate to severe DED from 27 United States centers. METHODS: Patients reported their medical history at baseline. They underwent ocular surface examinations and symptom evaluation using standardized protocols at baseline, 6 months, and 12 months. We analyzed the associations of systemic conditions (a systemic disease or smoking history) reported as potential DED risk factors with the severity of DED signs and symptoms using generalized linear regression models adjusted by age, gender, race, and visit. MAIN OUTCOME MEASURES: Dry eye disease symptoms assessed using the Ocular Surface Disease Index (OSDI), 6 DED signs (tear film break-up time, anesthetized Schirmer testing, corneal fluorescein staining, conjunctival lissamine green staining, tear osmolarity, and meibomian gland dysfunction), and a composite signs severity score from 0 to 1 (1 = most severe). RESULTS: The mean age was 58 years; 81% were women. More severe DED signs were associated significantly with Sjögren syndrome (mean composite signs severity score 0.52 with disease vs. 0.43 without disease; P < 0.001), facial rosacea (0.47 vs. 0.43; P = 0.002), rheumatoid arthritis (0.47 vs. 0.42; P = 0.002), peripheral artery disease (0.50 vs. 0.43; P < 0.001), and daily smoking history (0.45 vs. 0.43; P = 0.047). Thyroid dysfunction, osteoarthritis, diabetes, irritable bowel syndrome, hypercholesterolemia, hypertension, and hypertriglyceridemia were not associated significantly with DED signs. No conditions were associated significantly with OSDI. CONCLUSIONS: In this large, well-characterized cohort of patients with DED assessed under standardized procedures, patients with certain systemic diseases and smoking history showed more severe DED signs compared with patients without the conditions. The profile of significant DED signs varied by systemic condition, reflecting different DED causes. Understanding the systemic conditions and underlying causes that predispose some patients to severe DED can improve management.

Stability liabilities of biotherapeutic proteins: Early assessment as mitigation strategy.

Identification of molecular liabilities and implementation of mitigation strategies are key aspects that need to be considered by pharmaceutical companies developing therapeutic proteins. In the field of monoclonal antibodies, an efficient and streamlined process known as developability assessment is used for the selection of the "fittest" candidate. Other protein modalities, have in most cases only a limited number of possible candidates, requiring a paradigm change to a concept of candidate enabling. The assessment of liabilities at early project phases with the possibility to re-engineer candidates becomes essential for the success of these projects. Each protein possesses a unique stability profile resulting from the interplay of conformational, colloidal, chemical and physical stability attributes. All of these attributes strongly depend on external factors. Conformational and colloidal stability profiles of three non-immunoglobulin domain based proteins, namely Carbonic anhydrase, Ovalbumin and Thyroglobulin, and of two monoclonal antibodies were assessed in dependence of solution pH, ionic strength and varying buffering agents. The impact of screened external factors on proteins' stability attributes varied significantly, indicating presence of molecule specific liabilities. Screening of such a broad space of conditions at early project phases is only feasible using low-material consuming, high-throughput analytical methods as exemplified in this study.

Variability of urinary creatinine, specific gravity, and osmolality over the course of pregnancy: Implications in exposure assessment among pregnant women.

Due to dilution status of the urine, chemical concentrations measured in spot urine are frequently adjusted using correction factors, such as creatinine, specific gravity (SG), or osmolarity of the urine. Urinary correction factors, however, can be influenced dramatically by physiological changes such as pregnancy. Details about the variation of urine dilution over the course of pregnancy are not well characterized. In the present study, we investigated the variation of urine correction factors over time among the pregnant women of Korea (n = 69) and Thailand (n = 102). Creatinine, SG, and osmolality were determined in the urine samples obtained in each trimester of the participating women, and were compared by sampling time and by nationality. Implication of the variation in these correction factors was studied using phthalate metabolites measured in the urine samples as model chemicals. Urinary correction factors significantly varied across the trimesters especially in Korean pregnant women: urinary creatinine and osmolality were significantly lower in the third trimester (T3) urine than the first trimester (T1) urine. Urinary creatinine and SG of the T3 urine of Korean pregnant women were also significantly lower than those reported from the non-pregnant women who participated in Korean National Environmental Health Survey (KoNEHS) 2015-2017. Among Thai women, however, these correction factors were rather stable across the pregnancy. Differences in ethnicity, or in behavior such as water consumption amount may partly explain the differences. Temporal changes in these urine correction factors influenced the urinary phthalate metabolite concentrations adjusted for dilution, in both Korean and Thai pregnant women. The present observations show that the variations of urinary correction factors should be considered in exposure assessment of urinary chemicals for pregnant women, in order to circumvent potential bias due to physiological changes occurring during pregnancy, and to reduce errors in exposure classification and association.

Functional and Morphological Evaluation of Meibomian Glands in the Assessment of Meibomian Gland Dysfunction Subtype and Severity.

PURPOSE: To classify subtypes of meibomian gland dysfunction (MGD) and evaluate the dependency of dry eye signs, symptoms, and parameters on MGD subtype. DESIGN: Cross-sectional study. STUDY POPULATION: the right eyes of 447 patients with MGD of various subtypes and 20 healthy volunteers. METHODS: Patients were divided into 4 subtypes of MGD based on meibum expression, meibum quality, and MG loss on meibography images (meibograde of 0-6). Subtypes were patients with high meibum delivery (hypersecretory and nonobvious MGD) and those with low meibum delivery (hyposecretory and obstructive MGD). Additional clinical tests included tear film break-up time (TFBUT), ocular staining, osmolarity, Schirmer I, blink interval timing and the Ocular Surface Disease Index (OSDI) questionnaire. RESULTS: A total of 78 eyes had hypersecretory MGD; 49 eyes had nonobvious MGD; 66 eyes had hyposecretory MGD; and 254 eyes had obstructive MGD. Increased tear film osmolarity and lower TFBUT were found in the low-delivery groups; hyposecretory (P = 0.006, P = 0.016) and obstructive MGD (P = 0.008, P = 0.006) relative to high-delivery MGD (hypersecretory and nonobvious groups, respectively). Worse ocular symptoms and ocular staining were also found in low-delivery MGD groups than the high delivery MGD groups (P < 0.01 and P < 0.006, respectively). CONCLUSIONS: Patients with low-delivery MGD had worse dry eye parameters and ocular symptoms than those with high meibum delivery, indicating the pivotal role of meibum secretion in ocular surface health that should be targeted in MGD therapy. Furthermore, nonobvious MGD cannot be diagnosed using conventional dry eye tests and requires morphologic assessment of meibography images to confirm MG loss.

Afternoon urine osmolality is equivalent to 24 h for hydration assessment in healthy children.

BACKGROUND/OBJECTIVES: While daily hydration is best assessed in 24-h urine sample, spot sample is often used by health care professionals and researchers due to its practicality. However, urine output is subject to circadian variation, with urine being more concentrated in the morning. It has been demonstrated that afternoon spot urine samples are most likely to provide equivalent urine concentration to 24-h urine samples in adults. The aim of the present study was to examine whether urine osmolality (UOsm) assessed from a spot urine sample in specific time-windows was equivalent to 24-h UOsm in free-living healthy children. SUBJECTS/METHODS: Among 541 healthy children (age: 3-13 years, female: 45%, 77% non-Hispanic white, BMI:17.7 ± 4.0 kg m-2), UOsm at specific time-windows [morning (0600-1159), early afternoon (1200-1559), late afternoon (1600-1959), evening (2000-2359), overnight (2400-0559), and first morning] was compared with UOsm from the corresponding pooled 24-h urine sample using an equivalence test. RESULTS: Late afternoon (1600-1959) spot urine sample UOsm value was equivalent to the 24-h UOsm value in children (P < 0.05; mean difference: 62 mmol kg-1; 95% CI: 45-78 mmol kg-1). The overall diagnostic ability of urine osmolality assessed at late afternoon (1600-1959) to diagnose elevated urine osmolality on the 24-h sample was good for both cutoffs of 800 mmol kg-1 [area under the curve (AUC): 87.4%; sensitivity: 72.6%; specificity: 90.5%; threshold: 814 mmol kg-1] and 500 mmol kg-1 (AUC: 83.5%; sensitivity: 75.0%; specificity: 80.0%; threshold: 633 mmol kg-1). CONCLUSION: These data suggest that in free-living healthy children, 24-h urine concentration may be approximated from a late afternoon spot urine sample. This data will have practical implication for health care professionals and researchers.