Associations between fetal or infancy pet exposure and food allergies: The Japan Environment and Children's Study.

The hygiene hypothesis suggests that pet exposure is effective in preventing allergic disease, and some studies have reported the beneficial effects of dog exposure during fetal development or early infancy on food allergy. However, the effects of exposure to pets other than dogs on the kinds of food allergies remains unaddressed. This study aimed to explore the effect of exposure to various species of pets on the risk of food allergies. We obtained information on pet exposure and food allergy from the Japan Environment and Children's Study, a nationwide, prospective birth cohort study that included 97,413 mothers and their children. We examined the associations between exposure to various species of pets during fetal development or early infancy and the incidence risk of food allergies. We conducted logistic regression analysis for each pet species, causative food, and timing of exposure. Exposure to dogs or cats during fetal development or early infancy was estimated to reduce the incidence risk of food allergies until the age of 3 years. Dog exposure was estimated to reduce the incidence risk of egg, milk, and nut allergies, and cat exposure was estimated to reduce the incidence risk of egg, wheat, and soybean allergies. However, hamster exposure was estimated to increase the incidence risk of nut allergy. In conclusion, the association between pet exposure and food allergies might differ depending on the pet species and causative food. Continued dog and cat exposure from fetal development to infancy was estimated to reduce the incidence risk of food allergies. The findings of this study shall aid in the design of future studies.

Maternal prenatal and postnatal psychological distress trajectories and impact on cognitive development in 4-year-old children: the Japan Environment and Children's Study.

Maternal prenatal and postnatal psychological distress, including depression and anxiety, may affect children's cognitive development. However, the findings have been inconsistent. We aimed to use the dataset from the Japan Environment and Children's Study, a nationwide prospective birth cohort study, to examine this association. We evaluated the relationship between the maternal six-item version of the Kessler Psychological Distress Scale (K6) scores and cognitive development among children aged 4 years. K6 was administered twice during pregnancy (M-T1; first half of pregnancy, M-T2; second half of pregnancy) and 1 year postpartum (C-1y). Cognitive development was assessed by trained testers, using the Kyoto Scale of Psychological Development 2001. Multiple regression analysis was performed with the group with a K6 score ≤ 4 for both M-T1 and M-T2 and C-1y as a reference. Records from 1,630 boys and 1,657 girls were analyzed. In the group with K6 scores ≥ 5 in both M-T1 and M-T2 and C-1Y groups, boys had significantly lower developmental quotients (DQ) in the language-social developmental (L-S) area (partial regression coefficient: -4.09, 95% confidence interval: -6.88 - -1.31), while girls did not differ significantly in DQ for the L-S area. Among boys and girls, those with K6 scores ≤ 4 at any one or two periods during M-T1, M-T2, or C-1y did not have significantly lower DQ for the L-S area. Persistent maternal psychological distress from the first half of pregnancy to 1 year postpartum had a disadvantageous association with verbal cognitive development in boys, but not in girls aged 4 years.

Hormone-sensitive lipase protects adipose triglyceride lipase-deficient mice from lethal lipotoxic cardiomyopathy.

Lipid droplets (LDs) are multifunctional organelles that regulate energy storage and cellular homeostasis. The first step of triacylglycerol hydrolysis in LDs is catalyzed by adipose triglyceride lipase (ATGL), deficiency of which results in lethal cardiac steatosis. Although hormone-sensitive lipase (HSL) functions as a diacylglycerol lipase in the heart, we hypothesized that activation of HSL might compensate for ATGL deficiency. To test this hypothesis, we crossed ATGL-KO (AKO) mice and cardiac-specific HSL-overexpressing mice (cHSL) to establish homozygous AKO mice and AKO mice with cardiac-specific HSL overexpression (AKO+cHSL). We found that cardiac triacylglycerol content was 160-fold higher in AKO relative to Wt mice, whereas that of AKO+cHSL mice was comparable to the latter. In addition, AKO cardiac tissues exhibited reduced mRNA expression of PPARα-regulated genes and upregulation of genes involved in inflammation, fibrosis, and cardiac stress. In contrast, AKO+cHSL cardiac tissues exhibited expression levels similar to those observed in Wt mice. AKO cardiac tissues also exhibited macrophage infiltration, apoptosis, interstitial fibrosis, impaired systolic function, and marked increases in ceramide and diacylglycerol contents, whereas no such pathological alterations were observed in AKO+cHSL tissues. Furthermore, electron microscopy revealed considerable LDs, damaged mitochondria, and disrupted intercalated discs in AKO cardiomyocytes, none of which were noted in AKO+cHSL cardiomyocytes. Importantly, the life span of AKO+cHSL mice was comparable to that of Wt mice. HSL overexpression normalizes lipotoxic cardiomyopathy in AKO mice and the findings highlight the applicability of cardiac HSL activation as a therapeutic strategy for ATGL deficiency-associated lipotoxic cardiomyopathies.

Comparative effects of insulin glulisine and lispro on postprandial plasma glucose and lipid profile in Japanese patients with type 2 diabetes mellitus.

OBJECTIVE: The control of postprandial plasma glucose (PPG) excursions is critical in the prevention of diabetic complications. Controversy remains on the differences in postprandial actions of insulin glulisine and lispro. The aim of this study was to define the differences in the efficacy of these two insulin analogues on PPG. METHODS: The study subjects were 20 in-hospital patients with type 2 diabetes mellitus (T2DM). Plasma glucose (PG) was tightly controlled with basal insulin and insulin glulisine or lispro, and then glulisine or lispro were switched to the other insulin analog every other day for 6 study days. PG was measured before breakfast and 0.5-, 1-, and 2 h-postprandial during the study. Postprandial plasma C-peptide and lipids were analyzed in the first 2 days of the study. Postprandial increments in each parameter were compared between glulisine and lispro. RESULTS: Whereas the median value of 0.5 h-Δ-PPG was comparable in glulisine and lispro, the 1 h-Δ-PPG was significantly lower with lispro than with glulisine (41 vs 53 mg/dl, respectively, p = 0.03). Similarly, the 2 h-Δ-PPG with lispro was 10 mg/dl lower than that with glulisine (35 vs 45 mg/dl, respectively, p = 0.05). In parallel with PPG, Δ-C-peptide at 1- and 2 h-postprandial were significantly lower with lispro than glulisine (0.50 vs 0.75 ng/ml, respectively, and 0.55 vs 0.75 ng/ml, respectively). The increment in LDL-C and HDL-C was significantly lower with lispro than with glulisine at 0.5 h-postprandial. CONCLUSION: Insulin lispro seems superior to glulisine in the control of PPG in Japanese patients with T2DM.

Neonatal reticulocyte count during the early postnatal period.

BACKGROUND: There are few reports on reticulocyte count during the early postnatal period, and its clinical significance is not well understood. To examine the relationships between neonatal reticulocyte count and other perinatal variables. METHODS: We conducted a retrospective cohort study of neonatal infants who were admitted to the neonatal intensive care unit (NICU) of Ohta Nishinouchi Hospital, Japan, between April 1, 2016 and July 31, 2019. All blood samples were collected within 3 h after admission. RESULTS: Four hundred and twenty-eight infants were included, of whom 317 (74.1%) were preterm and 111 (25.9%) were term. Two hundred and ninety-nine infants (69.9%) were born by cesarean section. The median reticulocyte counts (‰) for all gestational ages (GAs) were as follows: 24-25 wks (n = 11), 124.1 (range: 106.3 to 148.6); 26-27 wks (n = 25), 111.1 (range: 55.5 to 144.3); 28-30 wks (n = 52), 81.9 (range: 35.6 to 131.5); 31-33 wks (n = 86), 71.6 (range: 28.3 to 116.6); 34-36 wks (n = 143); 59.6 (range: 30.2 to 110.9); and 37-41 wks (n = 111), 43.2 (range: 21.9 to 69.2). There were significant relationships between the neonatal reticulocyte count and gender [p < 0.01, odds ratio (OR), 0.37; 95% confidence interval (CI), 0.21 to 0.64], GA (p < 0.01, OR, 0.92; 95% CI, 0.90 to 0.93), delivery type (p = 0.03, OR, 0.51; 95% CI, 0.28 to 0.95), maternal haemoglobin before delivery (p < 0.01, OR, 0.74; 95% CI, 0.60 to 0.91), tracheal intubation at resuscitation (p = 0.04, OR, 2.75; 95% CI, 1.04 to 7.32) and mean platelet volume (p < 0.01, OR, 0.51; 95% CI, 0.35 to 0.74). CONCLUSION: A higher neonatal reticulocyte count in NICU infants may be one of the physiological responses to a more rapid environmental change during the early postnatal period.

Cardiac overexpression of perilipin 2 induces atrial steatosis, connexin 43 remodeling, and atrial fibrillation in aged mice.

Atrial fibrillation (AF) is prevalent in patients with obesity and diabetes, and such patients often exhibit cardiac steatosis. Since the role of cardiac steatosis per se in the induction of AF has not been elucidated, the present study was designed to explore the relation between cardiac steatosis and AF. Transgenic (Tg) mice with cardiac-specific overexpression of perilipin 2 (PLIN2) were housed in the laboratory for more than 12 mo before the study. Electron microscopy of the atria of PLIN2-Tg mice showed accumulation of small lipid droplets around mitochondrial chains, and five- to ninefold greater atrial triacylglycerol (TAG) content compared with wild-type (WT) mice. Electrocardiography showed significantly longer RR intervals in PLIN2-Tg mice than in WT mice. Transesophageal electrical burst pacing resulted in significantly higher prevalence of sustained (>5 min) AF (69%) in PLIN2-Tg mice than in WT mice (24%), although it was comparable in younger (4-mo-old) mice. Connexin 43 (Cx43), a gap junction protein, was localized at the intercalated disks in WT atria but was heterogeneously distributed on the lateral side of cardiomyocytes in PLIN2-Tg atria. Langendorff-perfused hearts using the optical mapping technique showed slower and heterogeneous impulse propagation in PLIN2-Tg atria compared with WT atria. Cardiac overexpression of hormone-sensitive lipase in PLIN2-Tg mice resulted in atrial TAG depletion and amelioration of AF susceptibility. The results suggest that PLIN2-induced steatosis is associated with Cx43 remodeling, impaired conduction propagation, and higher incidence of AF in aged mice. Therapies targeting cardiac steatosis could be potentially beneficial against AF in patients with obesity or diabetes.

Glycemic, insulinemic and incretin responses after oral trehalose ingestion in healthy subjects.

BACKGROUND: Trehalose is hydrolyzed by a specific intestinal brush-border disaccharidase (trehalase) into two glucose molecules. In animal studies, trehalose has been shown to prevent adipocyte hypertrophy and mitigate insulin resistance in mice fed a high-fat diet. Recently, we found that trehalose improved glucose tolerance in human subjects. However, the underlying metabolic responses after trehalose ingestion in humans are not well understood. Therefore, we examined the glycemic, insulinemic and incretin responses after trehalose ingestion in healthy Japanese volunteers. METHODS: In a crossover study, 20 fasted healthy volunteers consumed 25 g trehalose or glucose in 100 mL water. Blood samples were taken frequently over the following 3 h, and blood glucose, insulin, active gastric inhibitory polypeptide (GIP) and active glucagon-like peptide-1 (GLP-1) levels were measured. RESULTS: Trehalose ingestion did not evoke rapid increases in blood glucose levels, and had a lower stimulatory potency of insulin and active GIP secretion compared with glucose ingestion. Conversely, active GLP-1 showed higher levels from 45 to 180 min after trehalose ingestion as compared with glucose ingestion. Specifically, active GIP secretion, which induces fat accumulation, was markedly lower after trehalose ingestion. CONCLUSIONS: Our findings indicate that trehalose may be a useful saccharide for good health because of properties that do not stimulate rapid increases in blood glucose and excessive secretion of insulin and GIP promoting fat accumulation.

On the top of ARB N/L type Ca channel blocker leads to less elevation of aldosterone.

The activation of the renin-angiotensin system (RAS) is one of the unfavourable characteristics of calcium channel blocker (CCB). N type calcium channel is thought to be involved in renin gene transcription and adrenal aldosterone release. Accordingly, N/L type CCB has a possibility of less elevation of plasma aldosterone concentrations (PAC) among CCBs. In a monotherapy study, we had already demonstrated that N/L type CCB leads to less activation of the RAS compared with L type CCB. The objective of this study is to substantiate the hypothesis that at the condition of additive administration on the top of an angiotensin receptor blocker (ARB), still N/L type CCB leads to less elevation of PAC compared with L type one. Subjects were 60 hypertensives administered with valsartan. As an open label study, amlodipine (L type) or cilnidipine (N/L type) were administered on the top of valsartan (ARB) in a cross-over manner. Results were as follows (valsartan+amlodipine compared with valsartan+cilnidipine): systolic blood pressure (SBP)/diastolic blood pressure (DBP) (mmHg): 132±10/76±10 compared with 131±10/77±9, P=0.95/0.48, plasma renin activity (PRA) (ng/ml·h): 2.41±2.67 compared with 2.00±1.50 P=0.20, PAC (pg/ml): 77.3±31.0 compared with 67.4±24.8, P<0.05, urinary albumin excretion (UAE) (mg/gCr): 105.9±216.1 compared with 73.9±122.2, P<0.05. Thus, PAC at cilnidipine was significantly lower than those at amlodipine in spite of the comparable BP reductions. Besides, UAE was significantly lower at cilnidipine. In conclusion, on the top of the ARB, it is suggested that cilnidipine administration might lead to less elevation of PAC and reduction in UAE compared with amlodipine.

Daily Intake of Trehalose Is Effective in the Prevention of Lifestyle-Related Diseases in Individuals with Risk Factors for Metabolic Syndrome.

We previously performed animal studies that suggested that trehalose potentially prevents the development of metabolic syndrome in humans. To evaluate this possibility, we examined whether trehalose suppressed the progression of insulin resistance in a placebo-controlled, double-blind trial in 34 subjects with a body mass index (BMI) ≥23. The subjects were divided into two groups and were assigned to ingest either 10 g/d of trehalose or sucrose with meals for 12 wk. During the study, body composition and blood biochemical parameters were measured at week 0, 8, and 12. These parameters were also measured 4 wk after the end of intake to confirm the washout of test substances. In the trehalose group, blood glucose concentrations after a 2-h oral glucose tolerance test significantly decreased following 12 wk of intake in comparison with baseline values (0 wk). When a stratified analysis was performed in the subjects whose percentage of truncal fat approached the high end of the normal range, the change in body weight, waist circumference, and systolic blood pressure were significantly lower in the trehalose group than in the sucrose group. Our data indicated that a daily intake of 10 g of trehalose improved glucose tolerance and progress to insulin resistance. Furthermore, these results suggested that trehalose can potentially reduce the development of metabolic syndrome and associated lifestyle-related diseases, such as type 2 diabetes.

Determinants of plasma renin activity: role of a human renin gene variant as a genetic factor.

The plasma renin activity (PRA) is affected by a number of environmental factors. However, significant heritability has been shown for the activity. A hypothesis that a candidate regulatory single-nucleotide polymorphism, C-5312T, of human renin gene should have a significant effect on PRA was elucidated and updating of independent determinants of PRA was attempted. Cross sectional study. Outpatient study. We enrolled consecutive 810 subjects who had consulted our hospitals for lifestyle-related diseases. Genotypes were assayed with genomic DNA for C-5312T. Among the genetic variants, the difference of PRA was evaluated. Monovariate linear regression analysis was performed to test the correlation between PRA and clinical variables. Finally, stepwise multiple regression analysis was performed to evaluate the independent determinants. On comparing 2 genotype groups, CC/CT and T allele homozygote, the geometric means of PRA were 0.778 and 0.941 ng/ml/h, respectively (F = 5.992, P = 0.015). Monovariate linear regression analysis revealed that a number of variables have a significant correlation with the activity, including urinary salt excretion. A stepwise multivariate regression analysis revealed that renin C-5312T variant (TT) is one of the independent determinants of PRA. Thus, for the first time, a human renin gene variant was associated with a significant increase in PRA as a genetic factor and the independent determinants for the activity were updated including genetic factor.

Genetic variant of the renin-angiotensin system and prevalence of type 2 diabetes mellitus: a modest but significant effect of aldosterone synthase.

Recent genome-wide association studies have identified multiple variants that confer risk of type 2 diabetes mellitus (DM). However, established associations explain only a part of the heritability. Thus, even at the genome-wide association studies era, candidate gene approach should be still useful. Recent interventional studies against the renin-angiotensin system (RAS) showed reduction in new onset of DM, implying the system is involved in the onset. We substantiated the hypothesis that genetic variants of RAS have significant association with prevalence of DM. We enrolled to the study consecutive 782 subjects who had consulted our hospitals for mainly lifestyle related diseases. They consisted of 282 (36.1 %) diabetes cases. Genotypes were assayed with genomic DNA for conventional four genes of the RAS, i.e., angiotensin converting enzyme (ACE) insertion/deletion variant, angiotensinogen (AGT) M235T variant, angiotensin II type I receptor (AT1) A1166C variant, and aldosterone synthase (CYP11B2) C-344T variant. Association between the genetic variants of the RAS and prevalence of type 2 DM was tested. A significant association of DM and CYP11B2 genotype was obtained. There was no significant association between DM and ACE, AGT and AT1 variants. A multivariate logistic regression showed that age, gender, and CYP11B2 genotype were independent factors for association to diabetes, the DM risk of CC/CT to TT being 1.40 (95 % CI 1.04-1.90, p = 0.029). Thus, it is concluded that a genetic variant of the RAS should have a modest but significant impact on the onset of type 2 diabetes mellitus.

Trehalose prevents adipocyte hypertrophy and mitigates insulin resistance in mice with established obesity.

Our group recently demonstrated that simultaneous administration of trehalose with a high-fat diet (HFD) suppresses adipocyte hypertrophy and mitigates insulin resistance in mice. For the present study, we hypothesized that similar effects of trehalose would be observed in mice with previously-established obesity. Obese mice were fed a HFD and drinking water containing 0.3 or 2.5% (weight/volume) trehalose or distilled water (DW) ad libitum for 8 wk. After 7 wk intake of a HFD and trehalose, fasting serum insulin levels and homeostasis model assessment-insulin resistance (HOMA-IR) in the 0.3% Tre/HFD group were significantly lower than those in the DW/HFD group (p<0.05). After 8 wk of treatment, mesenteric adipocytes in the 0.3% Tre/HFD group showed significantly less hypertrophy than those in the DW/HFD group. Mechanistic analysis indicated that levels of high molecular weight (HMW) adiponectin in the serum of the 0.3% Tre/HFD group were significantly higher than those in the DW/HFD group. The expression levels of insulin receptor substrate-1 (IRS-1) and insulin receptor substrate-2 (IRS-2) messenger RNA (mRNA) in muscle were also significantly increased by trehalose intake. Our data therefore suggest that administration of trehalose to obese mice mitigates insulin resistance by suppressing adipocyte hypertrophy and increasing serum HMW adiponectin, resulting in upregulation of IRS-1, and IRS-2 expression in muscle. These results further suggest that trehalose is a functional saccharide that may be used to prevent the progression of insulin resistance.

Formation of a liquid organic ion associate in aqueous solution and its application to the GF-AAS determination of trace cadmium in environmental water as a complex with 2-(5-bromo-2-pyridylazo)-5-(N-propyl-N-sulfopropylamino)phenol.

The formation of a liquid organic ion associate in an aqueous sample was applied to the concentration and determination of cadmium in environmental water samples. Cadmium was converted into a complex with 2-(5-bromo-2-pyridylazo)-5-(N-propyl-N-sulfopropylamino)phenol (5-Br-PAPS) in a 40-mL sample solution, and was extracted into a liquid ion associate of phenolsulfonate and benzethonium during phase formation. More than 400-fold enrichment was easily attained by this technique, because the volume of the liquid organic phase formed was very small, ca. 2 microL. After dilution of the organic phase with a small volume of 2-methoxyethanol, the cadmium in the solution was determined by GF-AAS. The detection limit was 0.09 ng/L (3sigma(b)). This method was applied to the determination of cadmium in river water and seawater.

Metachromins R--T, new sesquiterpenoids from marine sponge Spongia sp.

Three new sesquiterpenoids, metachromins R--T (1--3), have been isolated from an Okinawan marine sponge Spongia sp. The structures and stereochemistry of 1--3 were elucidated on the basis of the spectroscopic data. Metachromins S (2) and T (3) showed modest cytotoxicity.

Suppression of apolipoprotein B secretion from HepG2 cells by glucosyl hesperidin.

Our previous study has shown that a soluble hesperidin derivative, glucosyl hesperidin (G-hesperidin), preferentially lowers serum triglyceride (TG) level in hypertriglyceridemic subjects through the improvement of very low-density lipoprotein (VLDL) metabolic abnormality. G-Hesperidin has also been found to decrease an elevated serum apolipoprotein B (apo B) level in the hypertriglyceridemic subjects, suggesting a possibility that this compound suppresses excess VLDL secretion in the liver. In the present study, to gain a better understanding of possible mechanisms by which G-hesperidin lowers serum TG, we examined whether this derivative affects apo B secretion from HepG2 human hepatoma cells, a model of hepatic VLDL secretion. As a result, G-hesperidin significantly reduced apo B secretion from the oleate-stimulated HepG2 cells. Furthermore, G-hesperidin significantly suppressed apo B secretion only in the oleate-stimulated cells and failed to act on the cells incubated without oleate. In the oleate-stimulated cells, G-hesperidin significantly decreased cellular cholesteryl ester (CE), although it had no effect on cellular TG or free cholesterol amounts. Moreover, the oleate-stimulated cells had a decrease in cellular apo B amounts by G-hesperidin exposure. These findings indicate that G-hesperidin down-regulates the assembly of apo B-containing lipoproteins via the reduction of CE synthesis augmented with oleate and results in suppressing excess apo B secretion from the cells. This effect is speculated to be associated with the improvement of VLDL metabolic abnormality in hypertriglyceridemic subjects and considered as a mechanism of lowering serum TG.

Bioavailability of glucosyl hesperidin in rats.

Glucosyl hesperidin (G-hesperidin) is a water-soluble derivative of hesperidin. We compared the absorption and metabolism of G-hesperidin with those of hesperidin in rats. After oral administration of G-hesperidin or hesperidin to rats, hesperetin was detected in sera hydrolyzed with beta-glucuronidase, but it was not detectable in unhydrolyzed sera. Serum hesperetin was found more rapidly in rats administered G-hesperidin than in those administered hesperidin. The area under the concentration-time curve for hesperetin in the sera of rats administered G-hesperidin was approximately 3.7-fold greater than that of rats administered hesperidin. In the urine of both administration groups, hesperetin and its glucuronide were found. Urinary excretion of metabolites was higher in rats administered G-hesperidin than in those administered hesperidin. These results indicate that G-hesperidin presents the same metabolic profile as hesperidin. Moreover, it was concluded that G-hesperidin is absorbed more rapidly and efficiently than hesperidin, because of its high water solubility.

Effects of free proline accumulation in petunias under drought stress.

Petunias (Petunia hybrida cv. 'Mitchell') accumulate free proline (Pro) under drought-stress conditions. It is therefore believed that Pro acts as an osmoprotectant in plants subjected to drought conditions. Petunia plants were transformed by Delta(1)-pyrroline-5-carboxylate synthetase genes (AtP5CS from Arabidopsis thaliana L. or OsP5CS from Oryza sativa L.). The transgenic plants accumulated Pro and their drought tolerance was tested. The Pro content amounted to 0.57-1.01% of the total amino acids in the transgenic plants, or 1.5-2.6 times that in wild-type plants grown under normal conditions. The transgenic plant lines tolerated 14 d of drought stress, which confirms that both P5CS transgenes had full functionality. Exogenous L-Pro treatment caused the plants to accumulate Pro; plants treated with 5 mM L-Pro accumulated up to 18 times more free Pro than untreated plants. Exogenous L-Pro restricted the growth of wild-type petunias more than that of Arabidopsis plants. The capacity for free Pro accumulation might depend on the plant species. The growth of petunia plants was influenced not only by the Pro concentration in the plants, but by the ratio of the Pro content to the total amino acids, because the growth of the transgenic petunia plants appeared normal.

Analysis of salt-stress-inducible ESTs isolated by PCR-subtraction in salt-tolerant rice.

To clarify the mechanisms of stress tolerance in rice and to search for rice genes associated with these mechanisms, we analyzed genes induced by a high salinity treatment using the PCR-subtractive hybridization method (PCR-subtraction). Seedlings of the salt-tolerant rice cultivar Dee-geo-woo-gen (DGWG) were either treated with 250 mM NaCl for 5 h or left untreated, and PCR-subtraction was then performed using the untreated (control) plants as a driver and the NaCl-treated plants as a tester. We obtained 384 clones of tester-specific cDNAs as salt-inducible candidates. Northern analysis performed with the cDNA fragments showed that 65 clones had been induced by the NaCl treatment. Sequence analysis and database searching indicated that these clones have homology to proteins functional for detoxification, stress response, and signal transduction in plants. Of these clones, 22% coded for unknown proteins and 12% gave no hits. We selected eight clones from each functional category and analyzed their expression pattern in DGWG. For temporal analysis, seedlings were treated with H(2)O or 250 mM NaCl for 0, 0.5, 1, 2, 5, 10 or 24 h. Different patterns of transcript regulation were found. For the analysis of expression in response to various types of stress and abscisic acid (ABA) treatments, seedlings were treated for 5 h or 10 h with H(2)O, dehydration, cold (4 degrees C), heat (40 degrees C), mannitol, ABA, or wounding. All clones were strongly up-regulated by osmotic stress (dehydration and mannitol) and the ABA treatment.