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Establishment of an AFP-producing serum-independent culture cell line of human liver cancer and changes in protein synthesis associated with the absence of serum (First Part).

We succeeded in the separation of AFP-producing cell lines (HH2-1 and HH2-6) and a non-AFP-producing cell line (HH2-4) from cultures of human hepatocellular carcinoma and also succeeded in culturing the AFP-producing cell lines in a serum-independent medium (HHSF). The cells cultured in this medium showed no morphological difference from those cultured in the medium containing serum, and retained growth ability well. Although the protein synthesis of the cells was reduced in the serum-independent medium, they were still able to produce a variety of proteins such as transferrin (TF) and fibronectin (FN) in addition to alpha-fetoprotein (AFP) and albumin (ALB).

Blood↗

Nutritional support treatment for severe chronic hepatitis and posthepatitic cirrhosis.

The therapeutic effectiveness of nutritional support in the treatment of severe chronic hepatitis and posthepatitic cirrhosis was evaluated. 143 patients with severe chronic hepatitis and 83 with posthepatitic cirrhosis were evaluated with SGA for assessing the nutritional status before the treatment. Patients with severe chronic hepatitis were divided into three groups: group A subject to enteral nutrition (EN) and parenteral nutrition (PN), group B subject to comprehensive treatment (CT)+PN; group C subject to CT+EN. The patients with posthepatitic cirrhosis were divided into two groups: group D receiving CT and group E receiving CT+PN+EN. The function of liver and kidney and nutritional status were monitored to assess the therapy in 6 weeks. The results showed before treatment, over 90 % patients had moderate to severe malnutrition. After nutritional support, the liver function (ALT, T-bil) and nutritional status (TP, TC) in group A was improved significantly as compared with that in groups B and C (P<0.05). Compared with group D, the values of TP and Alb were increased significantly in group E (P<0.05), but the levels of ALT, AST and T-bil had no obvious change. It was suggested that most patients with severe chronic hepatitis or posthepatitic cirrhosis had malnutrition to varying degrees. The nutritional support treatment could obviously improve the nutritional status of these patients, and was helpful to ameliorate the liver function of the patients with severe chronic hepatitis. Among the methods of nutritional support treatment, PN combined with EN had the best effectiveness.

Adolescent↗

Simple assay of serum copper fractions by ultrafiltration and flameless atomic absorption.

The authors present a simple and rapid method for assaying ultrafilterable copper (Cu UF) and albumin-bound copper (Cu ALB). It is based on the ultrafiltration of serum in the presence of ethylenediaminetetraacetic acid (EDTA), used to prevent adsorption on the membranes. EDTA at 0.4 g/L has no effect on the equilibrium of serum copper vectors and enables Cu UF to be assayed by flameless atomic absorption. EDTA at 2 g/L is used to assay total exchangeable copper (CU EXC) (ultrafilterable + albumin-bound). The evaluation criteria of the method are furnished, as are the normal values in healthy subjects: 14.6 micrograms/L for Cu UF and 87 micrograms/L for Cu EXC. Finally, the usefulness of the methods described here for the diagnosis of Wilson's and Menkes' disease was demonstrated.

Copper↗

Gene expression studies in prostate cancer tissue: which reference gene should be selected for normalization?

Using quantitative reverse transcription-polymerase chain reaction (RT-PCR), reference genes are utilized as endogenous controls for relative quantification of target genes in gene profiling studies. The suitability of housekeeping genes for that purpose in prostate cancer tissue has not been sufficiently investigated so far. The objective of this study was to select from a panel of 16 potential candidate reference genes the most stable genes for gene normalization. Expression of mRNA encoding ACTB, ALAS1, ALB, B2M, G6PD, GAPD, HMBS, HPRT1, K-ALPHA-1, POLR2A, PPIA, RPL13A, SDHA, TBP, UBC, and YWHAZ was examined in matched, microdissected malignant and nonmalignant tissue specimens obtained from 17 nontreated prostate carcinomas after radical prostatectomy by real-time RT-PCR. The genes studied displayed a wide expression range with cycle threshold values between 16 and 37. The expression was not different between samples from pT2 and pT3 tumors or between samples with Gleason scores <7 and >or=7 (P>0.05). ACTB, RPL13A, and HMBS showed significant differences (P<0.02 at least) in expressions between malignant and nonmalignant pairs. All other genes did not differ between the matched pairs, and the software programs geNorm and NormFinder were used to ascertain the most suitable reference genes from these candidates. HPRT1, ALAS1, and K-ALPHA-1 were calculated by both programs to be the most stable genes covering a broad range of expression. The expression of the target gene RECK normalized with HRPT1 alone and with the normalization factors generated by the combination of these three reference genes as well as with the unstable genes ACTB or RPL13A is given. That example shows the significance of using suitable reference genes to avoid erroneous normalizations in gene profiling studies for prostate cancer. The use of HPRT1 alone as a reference gene shown in our study was sufficient, but the normalization factors generated from two (HRPT1, ALAS1) or all three genes (HRPT1, ALAS1, K-ALPHA-1) should be considered for an improved reliability of normalization in gene profiling studies of prostate cancer.

Aged↗

Numerical and genetic analysis of polycyclic aromatic hydrocarbon-degrading mycobacteria.

Ability to degrade high molecular weight polycyclic aromatic hydrocarbons (PAHs) has been found in diverse species of fast-growing mycobacteria. This study included several PAH-degrading mycobacteria from heavily contaminated sites and an uncontaminated humus soil in the Natural Park, Schwäbische Alb, Germany. The numerical analysis with a total of 131 tests showed that isolates from humus soil and contaminated sites had similar substrate utilization patterns for primary alcohols from ethanol to pentanol, 1,4-butanediol, benzyl alcohol, hexadecane, ethyl acetate, fluoranthene, phenanthrene, and pyrene as the sole carbon and energy (C/E) sources. Significant differences between the two subgroups isolated from humus soil and contaminated sites were observed in the utilization of polyalcoholic sugars, including adonitol, D: -arabitol, L: -arabitol, erythritol, inositol, rhamnose, sorbitol, and xylitol. Among isolates from humus soil, strain PYR100 showed high similarity in 16S rDNA sequence with M. vanbaalenii strain PYR-1 (=DSM 7251, 100%) and M. austroafricanum ATCC 33464 (99.9%). In addition to the numerical analysis, the 16S-23S intergenic spacer sequence was useful for discriminating between the closely related strains PYR100 and PYR-1 (98% similarity). The patterns of the variable V2 and V3 regions in the ribosomal RNA gene corresponding to Escherichia coli positions 179 to 197 and 1006 to 1023, respectively, were useful for dividing fast-growing and thermosensitive PAH-degrading mycobacteria into ten subgroups consistent with the phylogenetic positions.

Base Sequence↗

Linkage between vitamin D-binding protein and alpha-fetoprotein in the mouse.

The albumin gene family consists of four evolutionarily related genes that code for serum transport proteins. In rodents, the genes for albumin, alpha-fetoprotein, and alpha ALB are physically linked within 100 kilobases of DNA. The fourth gene, Gc, encoding vitamin D-binding protein or group-specific component, maps to the same chromosome as the other family members, but linkage has not been established. This report describes the genetic and physical mapping of Gc in mouse and establishes that, although Gc is genetically linked to the other genes, its physical distance from them extends beyond the resolution range of yeast artificial chromosome cloning and pulsed-field gel electrophoresis.

Animals↗

Pigs with the dominant white coat color phenotype carry a duplication of the KIT gene encoding the mast/stem cell growth factor receptor.

Comparative mapping data suggested that the dominant white coat color in pigs may be due to a mutation in KIT which encodes the mast/stem cell growth factor receptor. We report here that dominant white pigs lack melanocytes in the skin, as would be anticipated for a KIT mutation. We found a complete association between the dominant white mutation and a duplication of the KIT gene, or part of it, in samples of unrelated pigs representing six different breeds. The duplication was revealed by single strand conformation polymorphism (SSCP) analysis and subsequent sequence analysis showing that white pigs transmitted two nonallelic KIT sequences. Quantitative Southern blot and quantitative PCR analysis, as well as fluorescence in situ hybridization (FISH) analysis, confirmed the presence of a gene duplication in white pigs. FISH analyses showed that KIT and the very closely linked gene encoding the platelet-derived growth factor receptor (PDGFRA) are both located on the short arm of Chromosome (Chr) 8 at band 8p12. The result revealed an extremely low rate of recombination in the centromeric region of this chromosome, since the closely linked (0.5 cM) serum albumin (ALB) locus has previously been in situ mapped to the long arm (8q12). Pig Chr 8 shares extensive conserved synteny with human Chr 4, but the gene order is rearranged.

Animals↗

Urinary protein excretion in humans exposed to arsenic and cadmium.

OBJECTIVE: In mice, the renal toxicity of arsenic (As) and cadmium (Cd) has been shown to be exacerbated by the simultaneous administration of both elements. To verify the existence of such an interaction in humans, cohorts slightly (Belgian) and moderately (Chinese) exposed to both elements were examined. METHODS: Biological indicators of exposure (Cd in urine and in blood; As in urine) and renal effect parameters (retinol binding protein (RBP); beta(2)-microglobulin (beta(2)M); albumin (ALB); N-acetyl-beta-glucosaminidase (NAG) in urine) were determined and their relationships studied by multiple regression analyses. RESULTS: Changes in renal effect parameters could be ascribed to Cd body burden. RBP and beta(2)M urinary concentration were influenced by exposure to As only in Chinese women, directly and in interaction with Cd exposure. CONCLUSION: A synergistic action of As on the tubular effects of Cd is observed in women moderately exposed to these elements and leads to RBP urinary excretion slightly above normal values.

Acetylglucosaminidase↗

Postoperative evaluation of pylorus-preserving procedures compared with conventional distal gastrectomy for early gastric cancer.

We evaluated postoperative function in 98 patients who underwent surgery for early gastric cancer between 1995 and 1998 to compare the results of pylorus-preserving procedures to those of conventional distal gastrectomy with Billroth I (B-I). The pylorus-preserving procedures included endoscopic mucosal resection (EMR), performed in 12 patients; local resection (Local), performed in 14 patients; segmental resection (Seg), performed in 8 patients; and pylorus-preserving gastrectomy (PPG), performed in 19 patients. B-I was performed in 45 patients. The nutritional status and serum albumin (Alb) levels after PPG, the hemoglobin (Hb) levels after EMR, Local, and PPG, and the present/preoperative body weight ratios after EMR, Local, Seg, and PPG were superior to those after B-I. The time before oral intake was recommenced after EMR and Local, the volume of oral intake tolerated after EMR, Local, Seg, and PPG, and the postoperative hospital stay after EMR were all superior to those after B-I. Moreover, significantly fewer patients suffered reflux symptoms after EMR, Local, and PPG, abdominal fullness after EMR, and early dumping syndrome after EMR, Local, and PPG than after B-I. There was also less evidence of gastritis after EMR, Local, and PPG, and of bile reflux after EMR, Local, and PPG, than after B-I. These findings indicate that pylorus-preserving procedures may result in a better postoperative quality of life for selected patients with early gastric cancer.

Adult↗

Unravelling the biological nexus of smoking and postpartum depression: a meta-analysis and functional genomics approach.

PURPOSE: Postpartum depression (PPD) is a prevalent psychological condition among birthing women. While several psycho-socio-economic and neurobiological factors influence its development, its relationship with smoking behavior and nicotine addiction remains largely inconclusive. METHODS: In this combinatorial study, we first evaluate the relationship between smoking and depressive behaviors in postpartum women using data extracted from pertinent primary epidemiological studies. Additionally, to discern the molecular and cellular mechanisms underlying this association, we identified common genetic elements and evaluated their functional attributes using in silico analyses. RESULTS: Meta-analytical assessment of systematically collected data from 38 studies indicated that smoking women are twice as likely to develop PPD, compared to their non-smoking counterparts. While geocultural attributes did not affect this relationship, timing of smoking was a significant moderator, with current and gestational smoking statuses being more strongly linked with PPD outcome, compared to the past smoking habit. Further, depression scores in smoking postpartum women were higher than those in non-smoking controls. Analysis of the common protein-encoding genes underlying the pathophysiology of nicotine addiction and PPD revealed several critical hub proteins (viz., AKT1, JUN, CTNNB1, PTEN, EGFR, ESR1, SRC, STAT3, FN1, IL1B, IL6, TNF, TP53, GAPDH, INS, MYC, and ALB) which were predicted to alter multiple pathophysiological pathways associated with transcriptional expression, intra- and intercellular signaling transduction, metabolism, and immune functions. CONCLUSION: Our results indicate that smoking is strongly associated with depressive behavior in postpartum women, although this association involve mediation of additional environmental and psychosocial elements. Moreover, network analysis of common genetic elements identified several potentially disrupted neurophysiological pathways in postpartum women with smoking and depressive behaviors which may aid in characterizing the underlying relationship between the two conditions.

Humans↗

Comparison of various biochemical measurements with bone mineral densitometry and quantitative ultrasound for the assessment of vertebral fracture.

We have retrospectively studied postmenopausal elderly Japanese women (n = 288; age range, 60-75 years, 65.8 +/- 4.5 [mean +/- SD]) for the evaluation of biochemical measurements in assessment of bone mass and vertebral fracture, comparing with several bone mineral measurements and quantitative ultrasound (QUS) measurement. Several biochemical parameters [red cell count (RBC), hemoglobin (HB), albumin (ALB), and cholesterol (CHO)] positively correlated with bone mass parameters, but only serum cholesterol showed association with the presence of vertebral fracture. Urinary deoxypyridinoline (DPYD) and n-telopeptide (NTx) showed moderate negative correlation with bone mass parameters, and DPYD showed association with vertebral fracture. All bone mineral measurements (lumbar spine, total body, femoral neck by DXA, calcaneal bone by SXA, distal radius by pQCT) and QUS measurement (os calcaneus by two different QUS machines) showed a higher odds ratio and high chi2 value in logistic regression analysis for association with vertebral fracture. Thus, bone mass measurement is the principal method for assessment of fracture risk, and biochemical measurement should be used for motivation of further bone mass measurement. In biochemical measurements, measurement of serum cholesterol is cheap and easy, and thus might have an advantage, although further study is necessary.

Absorptiometry, Photon↗

Identification and analysis of metabolic reprogramming-related genes in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is notorious for its rapid progression, tendency to metastasize, high recurrence rates, dismal outcomes, and limited treatment options, underscoring the urgent need to uncover new biomarkers and molecular pathways to enhance diagnosis, prognosis, and therapeutic strategies. Metabolic reprogramming continues to play a role throughout the life cycle of cancer, evolving and adapting. In this study, we aimed to identify specific genes associated with metabolic reprogramming in TNBC, which can potentially become unique biomarkers of this cancer. TNBC datasets retrieved from the Gene Expression Omnibus were employed to pinpoint genes exhibiting altered expression linked to tumor metabolic reprogramming. Key genes were accurately screened through machine learning algorithms, and then externally verified using the TBNC dataset based on the Cancer Genome Atlas database. Finally, immunohistochemical methods were used to clinically confirm the differential expression and trends of these key genes. Our analysis accurately identified four genes-CLEC7A, IRS1, RSPO3, and ALB-that are closely correlated with the metabolic reprogramming characteristics of cancer, and could be regarded as innovative biomarkers for TNBC. This opens a new avenue for further investigation into the mechanisms of metabolic reprogramming in TNBC and new treatment strategies.

Humans↗

Reproductive factors and breast cancer risk among older women.

Reproductive factors have been shown to affect pre- and postmenopausal breast cancer risk differently, but whether there are additional age-specific differences among menopausal women as they age has not been clarified. We analyzed data from a large population-based case-control study that included 1,508 breast cancer cases and 1,556 controls, aged 20-98 years, who completed an in-home interviewer-administered questionnaire. The following subgroups were created to examine if the associations between reproductive factors and breast cancer risk varied by age- and menopausal-status: premenopausal (n=968), postmenopausal <65 years (n=1,045), postmenopausal >or=65 years (n=958). Among postmenopausal women >or=65 years, ever having breastfed decreased risk (odds ratio (OR)=0.67, 95% confidence interval (CI)=0.48, 0.92), and a strong dose-response relationship was observed for longer durations of breastfeeding (P trend=0.02), with the most pronounced protective effect observed for >or=14 months of breastfeeding (OR=0.40, 95% CI=0.21,0.76). Late age at first birth (AFB) and older age at last birth (ALB) were associated with non-statistically significant increases in breast cancer risk in this older group, while late age at menarche and surgical menopause decreased risk. ORs for multiparity were close to the null. Among premenopausal women and postmenopausal women <65 years, multiparity significantly decreased risk, and older AFB nonsignificantly increased risk. Our findings suggest that the well-known protective effect of multiparity attenuates with older age. Moreover, breastfeeding, one of the few potentially modifiable risk factors for breast cancer, was an important factor in decreasing risk among older parous postmenopausal women.

Adult↗

Associated factors in modulating aflatoxin B1-albumin adduct level in three Chinese populations.

To elucidate the potential factors modulating exposure to aflatoxin B1 (AFB1) in three Chinese populations, an epidemiologic study was conducted in Fusui County and Nanning City of Guangxi Province and Chengdu City of Sichuan Province. The incidence rates of hepatocelluar carcinoma (HCC) for males in these three regions were 92-97 per 100,000, 32-47 per 100,000, and 21 per 100,000, respectively. Eighty-nine residents from Fusui, 196 residents from Nanning, and 118 residents from Chengdu were screened for AFB1-albumin adduct (AAA) levels and hepatitis virus (HBV, HCV, HDV, HEV, and HGV) infections, as well as liver biochemistry (alanine aminotransferase [ALT], aspartate aminotransferase [AST], alkaline phosphatase [ALP], y-glutamyl transpeptidase [GGT], 5'-nucleotidase, globulin [GLO], direct bilirubin, indirect bilirubin, and bile acid levels). At least one marker of hepatitis virus (HV) infection was present in 47.2% (42/89) of subjects from Fusui, while in Nanning and Chengdu the values were 15.8% (31/196) and 22.0% (26/118), respectively. In contrast to females, a higher level of AAA was observed in males; the difference was statistically significant in both the Nanning (P = 0.023) and the Chengdu (P = 0.026) subjects. In the Chengdu group, there was a significantly higher level of AAA in cases with HV infection (P = 0.041). There was a close association between AAA level and BMI in the adults without HV infection (r = 0.148, P = 0.044). Also, AAA was closely associated with DBIL and GGT in non-HV-infected minors (P < 0.05), closely associated with ALB, GLO, and GGT in HV-infected minors (P < 0.05), and closely associated with IBIL, GLO, TBA, and AST in non-HV-infected adults (P < 0.01). The co-effect of HV infection and AFB1 exposure may be responsible for the high risk of HCC in the Fusui region, whereas age, gender, BMI, and HV infection may modify individual aflatoxin levels. The relationship between AAA level and liver biochemistry indicates injury induced by aflatoxin to both hepatic parenchyma and biliary tract. But the associations vary with age and HV infection status.

Aflatoxin B1↗

Vitamin E protects against oxidative damage caused by formaldehyde in the liver and plasma of rats.

It is well known that formaldehyde (FA) and reactive oxygen species (ROS) are cytotoxic and potentially carcinogenic. Although the individual effects of these reactants on cells have been investigated, the cytotoxicity exerted by the coexistence of FA and ROS is poorly understood. The present study was carried out to evaluate oxidant/antioxidant status and biochemical changes occurring after chronic formaldehyde toxicity in liver tissue and plasma of rats and protective effect of vitamin E (vit E) against oxidative damage. Eighteen rats were divided into three groups: (1) control rats, (2) rats treated with FA (FAt), and (3) rats treated with FA plus vit E (FAt + vit E) groups. After the treatment, the animals were sacrificed and liver tissues were removed for biochemical investigations. As a result, FA treatment significantly increased the levels of tissue malondialdehyde (MDA), protein carbonyl (PC), nitric oxide (NO) and the activity of xanthine oxidase enzyme (XO). On the other hand, FA exposure led to decrease in superoxide dismutase (SOD) and catalase (CAT) activities in liver tissues compared to control. FA caused significant decreases in total protein (TP) and albumin (ALB) whereas increases in aspartate transaminase (AST), alanine aminotransferase (AST), alkaline phosphatase (ALP) and interleukine-2 (IL-2) levels in plasma. Vit E treatment abolished these changes at a level similar to the control group. It was concluded that vit E treatment might be beneficial in preventing FA-induced liver tissue damage, and therefore have potential for clinical use.

Animals↗

In vitro differentiation of mouse ES cells into hepatocytes with coagulation factors VIII and IX expression profiles.

Coagulation factors II, V, VII, VIII, IX and X are produced by hepatocytes. So factors VIII and IX deficiencies, which result in hemophilia A and B, have the potential to respond to cellular replacement therapy. Embryonic stem (ES) cells provide a unique source for therapeutic applications. Here, E14 mouse ES cells have been induced into hepatocytes in vitro. Morphology revealed that ES-derived hepatic-like cells were round or polyhedral shaped with distinct boundary of individual cells, and some arranged in trabeculae. These cells expressed endodermal- or liver-specific mRNA--transthyretin (TTR), alpha1-anti-trypsin (AAT), alpha-fetoprotein (AFP), albumin (ALB), glucose-6-phoshpatase (G6P) and tyrosine aminotransferase (TAT). Approximately (85.1 +/- 0.5)% of the ES-derived cells was stained positive green with ICG uptake. These cells were also stained magenta as a result of PAS reaction. In this paper, expression of coagulation factors VIII and IX mRNA in the ES-derived cells is documented. Therefore, ES cells might be developed as substitute donor cells for the therapy of coagulation factor deficiencies.

Animals↗

The relationship between serum levels of lipoprotein(a) and proteins associated with the acute phase response.

The association of serum lipoprotein(a) (Lp(a)) with inflammation was investigated in a primarily rheumatologic study group (n = 570; 202 males and 368 females) by studying the relationship between serum levels of Lp(a) and a panel of acute phase proteins (C-reactive protein (CRP), alpha 1-antitrypsin (AAT), alpha 1-acid glycoprotein (AGP), haptoglobin (HPT), complement components 3 and 4 (C3, C4), prealbumin (PAL), albumin (ALB) and transferrin (TRF)). Lp(a) data were adjusted for age and sex, but not clinical condition as no significant differences in Lp(a) levels were observed, using analysis of variance, among the 15 diagnostic categories in the study group. Univariate analyses revealed significant positive associations between Lp(a) levels and levels of C4, AGP, C3 and HPT. Multivariate analysis revealed that C4 and AGP (in descending order of significance) were significant independent predictors of Lp(a) concentration, together accounting for 2.9% of the variability in Lp(a) concentration in the present study group. The data indicate that confounding effects of an acute phase response should be considered in epidemiologic studies, if a high prevalence of inflammation is suspected.

Acute-Phase Proteins↗

Correlations between EEG and chemical changes in uremia.

This study investigated correlations between the EEG and 10 different chemical indices in 23 uremic patients, tested over long periods of time (up to 18 months). An abnormal EEG was recorded at least once in 70% of these patients, but among the 362 tracings from all patients, 36% were considered abnormal. Slow wave abnormalities, mainly mild in degree and frontal in location, were found in 97% of abnormal records and epileptiform patterns in the form of bilateral spike and wave complexes were seen in 14%. Patients with these EEG abnormalities had a significantly slower frequency of background rhythm than patients with normal EEGs. Compared to those with normal records, patients with abnormal EEGs had a significantly higher BUN and Cl and lower Ca, as determined by standard (absolute) mean values. With relative changes, based on the percentage change from the previous value, the BUN, Ca, Cl, K, PO4 and CO2 showed significant relationships with a deteriorating EEG and the Ca, Cl, K and CO2 with an improving record. A deteriorating EEG was usually associated with an increasing BUN, Cr, K, Cl and Na and a decreasing Ca, PO4, Hct, Alb and CO2. Significant absolute changes in the chemical indices were more often seen than relative ones, especially with a deteriorating EEG. With either absolute or relatives values, the BUN showed the closest relationship with EEG deterioration and the Cr, Cl and Hct with EEG improvement. Changes were often seen in a combination of chemical indices, most often involving the BUN and to a lesser extent the Cr. With combined changes the one with the greatest change was especially the BUN and PO4. Variations in the amount of epileptiform activity were most closely related to changes in the BUN. The main conclusions of this study are that the BUN correlated best with the EEG among the 10 different serum values using various indices, especially standard (absolute) values with a deteriorating recording. The second highest number of correlations were found with the Cl, especially absolute values with an improving EEG. The Ca and K showed the best correlation with relative changes. Thus, correlations of various chemical indices with electrographic changes in uremic patients depend upon whether absolute or relative values are used and whether EEGs are deteriorating or improving.

Blood Urea Nitrogen↗