bluuebunny/crossref_metadata_2025 huggingface dataset:
Up‐regulation of Adrenal Leptin Receptor Expression and Tyrosine Hydroxylase Activation in Age‐related Hypertension
Authors: Green Jessica , Shek Eugene W
Published: June 2021

Blood pressure (BP) and sympathetic activation have been shown to increase with age. The aim of the present study was to examine whether increased adrenal OB‐Rb expression and tyrosine hydroxylase (TH) activation mediated age‐related hypertension.

Method

4‐6 mth (young) and 20‐24 mth (old) male F344/BN rats (n=4‐6 rats/age) were implanted with telemetry for 24‐hrs arterial pressure (MAP) monitoring and were housed in individual metabolic cages. Adrenal leptin receptors (OB‐Ra & OB‐Rb) and TH protein expressions were determined by Western Blot.

Results

Old rats exhibited a significant increase in MAP (107±1 vs 98±1mmHg), body weight (556±11 vs 332±5g) and serum leptin (24±5 vs 5.1±0.7ng/ml) compared with young rats (p<0.05). The increased MAP was positively correlated with serum leptin (r2=0.7, p<0.0001). Western Blot analysis revealed a higher OB‐Rb expression (54%) than OB‐Ra in young adrenal. Although OB‐Ra expression did not change with age, OB‐Rb protein expression was markedly up‐regulated by 211±13% in old compared with young rats. TH activation (as measured by phosphorylated TH/total TH) in adrenal was also elevated by 185±10% in old compared with young rats.

Conclusion

Results of the present study suggest that the increased BP with age may, at least in part, be mediated by the elevated circulating leptin levels, up‐regulated adrenal OB‐Rb protein expression and TH activation.
Pericyte‐mediated regulation of vasa recta capillaries in situ.
Authors: Peppiatt‐Wildman Claire M , Crawford Carol , Desai Tanmay D , Unwin Robert J
Published: June 2021

Pericytes have been proposed as functional regulators of capillary diameter in situ and thus blood flow in the CNS [1]. Pericytes occur at regular intervals on renal medullary vasa recta capillaries and are known to constrict isolated perfused descending vasa recta [2]. In this study we show for the first time pericyte‐mediated constriction of vasa recta in situ, in live kidney slices.

Angiotensin‐II (Ang‐II) is a potent vasoconstrictor in the kidney. Ang‐II (100 nM) evoked significant vasoconstriction of vasa recta at pericyte sites (41.1+6.2%) versus non‐pericyte sites (9.9+5.2%). Noradrenaline (NA, 10‐100 µM) also evoked a significantly greater vasoconstriction of vasa recta at pericyte sites (22.1+3.3%) than at non‐pericyte sites (9.2+1.8%). ATP (10 nM‐10 µM), which is also a vasoconstrictor agent endogenous to the kidney failed to evoke significant constriction of the vasa recta. The NO donor SNAP, evoked a significantly greater vasodilation of vasa recta at pericyte sites (18.6+5.3%) than that at non‐pericyte sites (4.7+2.8%). Data presented here indicates that in the kidney, like in the CNS, pericytes constrict vasa recta vessels and may play a role in regulation of blood flow in the renal medulla. Funded by the MRC.

[1] Peppiatt CM, Howarth C, Mobbs P, Attwell D. Nature. 2006 Oct 12;443(7112):700‐4

[2] Zhang Z, Rhinehart K, Kwon W, Weinman E, Pallone TL. Am J Physiol Heart Circ Physiol. 2004 Aug;287(2):H773‐81
Aquaporin‐1 is expressed in the thick ascending limb.
Authors: Herrera Marcela
Published: June 2021

The thick ascending limb (TAL) is the water‐impermeable segment of the nephron. Isolated TALs exhibit low luminal water permeability even in the presence of AVP, a characteristic that allows this segment to dilute the forming urine. Thus it is assumed that TALs do not express water channels. However, TALs rapidly shrink in response to increased basolateral osmolality suggesting the presence of water channels. We hypothesized that the TAL expresses AQP1. We measured AQP1 by: 1) Western blot, 2) RTPCR, and 3) immunofluorescence. By Western blot, homogenates of rat TAL suspensions exhibited two bands characteristic of glycosylated (0.8 ± 0.5 OD units) and unglycosylated (3.4 ± 0.5 OD units) AQP1 (n=5). To ensure that the signal was not due to other cell types in our preparation, we used microdissected TALs. AQP1 was detected in microdisected rat and mice TALs. No bands were detected in TALs from AQP1 ‐/‐ mice. By RTPCR, microdissected rat TALs, showed the predicted 430 bp and 603 bp products for AQP1 and the NaK2Cl co‐transporter (specific for TALs). Double immunofluorescence labeling showed both, basolateral localization of AQP1 and luminal localization for Tham Horsfall protein (specific for TALs). We conclude that AQP1 is expressed in the basolateral side of the TAL. Since the TAL is water impermeable, these data suggest AQP1 could be involved in transport of molecules other than water in this nephron segment.
Studies on gene expression in the kidneys of P2Y2 receptor knockout mice
Authors: Tuttle Jonathon D. , Kishore Bellamkonda K.
Published: June 2021

Research conducted in the host laboratory showed that P2Y2 receptor (P2Y2‐R) plays a key role in the maintenance of water homeostasis in the renal medullary collecting duct by interacting with vasopressin and prostanoid systems. Based on these observations, the aim of this study is to examine the gene expression of key molecules of prostanoid biosynthesis and NOS isoforms in P2Y2‐R knockout (KO) and wild type (WT) mice. RNA was extracted from the cortex and medullary tissue samples of KO and WT mice (N = 5 per group). Reverse transcription (RT) was performed on the RNA samples using SuperScript III Super Mix. Quantitative PCR was performed on the cDNA samples using gene specific primers for COX‐1, COX‐2, mPEGS‐1, nNOS, iNOS, eNOS and β‐actin and TaKaRa Taq R‐PCR Version in the Smart Cycler II System. SYBR Green was used for detection. Relative expression values were computed by normalizing the expression of target genes to that of β‐actin. Statistical analysis showed that expression of none of the molecules studied was significantly altered in the KO mice as compared to WT mice, although, numerically higher or lower values were observed for some of the molecules. These data indicate that genetic deletion of P2Y2‐R may not significantly alter the expression of molecules of prostanoid biosynthetic pathway and NOS isoforms at least under basal conditions. It is possible that when challenged by subjecting the KO and WT mice to experimental conditions that are known to alter the prostanoid biosynthesis or NOS isoforms, significant differences between the KO and WT mice could be observed. Work supported by the Dept. of Veterans Affairs and NKF of Utah and Idaho.
Effect of type 1 diabetes on protein kinase C (PKC) in rat renal medullary thick ascending limb
Authors: YANG JING , Pollock Jennifer S. , Carmines Pamela K.
Published: June 2021

Type 1 diabetes (T1D) accelerates superoxide production by the medullary thick ascending limb (mTAL), and this phenomenon is markedly attenuated by PKC inhibition (FASEB J 21:578.6, 2007). We hypothesized that T1D increases PKC activity and the expression of one or more PKC isoforms in the mTAL. mTAL suspensions were prepared from rats with streptozotocin‐induced T1D (20‐25 days after onset; blood glucose = 383 ± 13 mg/dl) and from normal or Sham (vehicle‐treated) rats. Total PKC activity, measured with a commercially‐available kit, was higher in mTALs from T1D rats than those from normal rats (1.38 ± 0.06 vs 0.73 ± 0.05 O.D./μg protein; P < 0.05). Calphostin C (1 μM) significantly reduced PKC activity to similar values (~0.2 O.D./μg protein) in mTALs from both groups. Western blot analysis revealed elevated PKC‐α protein levels (normalized to β‐actin) in mTALs from rats with T1D (170 ± 19% of Sham; P < 0.05). PKC‐β protein levels were also modestly elevated in mTALs from T1D rats (129 ± 9% of Sham; P < 0.05). In contrast, PKC‐δ levels did not differ between groups (values from T1D rats averaged 105 ± 17% of Sham). These data confirm that T1D increases PKC activity in the mTAL, in association with upregulation of PKC‐α and PKC‐β (but not PKC‐δ). We speculate that the changes in PKC‐α and PKC‐β expression may contribute to increased PKC activity and, in turn, to accelerated superoxide production by the mTAL during T1D.
Diabetes alters the production and release of catecholamines in Primary Mesangial Cell Culture
Authors: Cunha Tatiana S , Arita Danielle Yuri , Nogueira Marie Doki , Perez Juliana Dinéia , Andrade Maria Claudina Camargo , Casarini Dulce Elena
Published: June 2021

Catecholamines (CA) are produced by mesangial cells (MC), and their levels are increased in diabetes. The aim was to asses the effect of diabetes duration on the production/release of CA‐dopamine (DA), epinephrine (EP) and norepinephrine (NE)‐in cultured MC. Male Wistar rats were randomized into Control, Diabetes 3M (3 months diabetes) and Diabetes 4M (4 months diabetes) groups. Rats were made diabetetic (streptozotocin, 50mg/Kg) and after 3 or 4 months, kidneys were excised for glomeruli isolation. MC were cultured in DMEM, CA were extracted and quantified by high performance liquid chromatography. Extracellular level of EP was increased in Diabetes 3M group, as compared to control group (55±9 vs. 29±3 pg/mg protein). On the other hand, 4 months of diabetes increased intracelular concentration of EP (77±10 pg/mg protein) and NE (76.5±10 pg/mg protein) compared with control and Diabetes 3M groups, with no effect on DA levels. Intracellular concentration of EP and NE was not affected after 3 months of diabetes; however DA level was significantly decreased in Diabetes 3M group (81±5 pg/mg), as compared to control (under detection limit). In Diabetes 4M group, we observed an increase in intracellular concentration of EP (151±27 pg/mg protein) and NE (53±14 pg/mg protein) and a decrease of DA levels. Our results show that diabetes duration influences the production/release of catecholamines by MC and suggests that this pathway can have a role on renal dysfunction induced by time‐duration of diabetes.

Financial Support: CNPq.
Effects of triclosan on the normal intestinal microbiota and on susceptibility to experimental murine colitis
Authors: Pasch Erica , Voltmer Leah , Gemmell Summer , Walter Jens , Walton Kristen L.W.
Published: June 2021

Evidence suggests that alterations in the intestinal microbiota may be a factor in symptoms of inflammatory bowel disease. Triclosan, an antibacterial agent found in consumer products, is detectable in wastewater and in urine from sample human populations. The current study determined whether low level exposure to triclosan, mimicking environmental exposure, alters intestinal bacterial populations and susceptibility to colitis. Mice were given triclosan in the drinking water (10 μg/ml) for 5 days, followed by induction of colitis by 3% dextran sulfate sodium (DSS). Fecal bacteria were cultured to determine CFUs of total anaerobes, lactobacilli, and coliforms. Fecal DNA was analyzed by PCR and denaturing gradient gel electrophoresis (DGGE) using universal primers targeting the 16S rRNA gene. Colitis was assessed by disease activity index, histological scoring, and RT‐PCR for cyclooxygenase‐2. Clinical colitis scores were increased with DSS but were not altered by triclosan treatment. Total anaerobes were increased by DSS but were not altered by triclosan, while lactobacilli were decreased by triclosan and DSS. DGGE analysis indicated a shift in the fecal bacterial population with DSS that was altered by triclosan + DSS. We conclude that low‐level exposure to triclosan does not significantly alter susceptibility to DSS colitis, but that it may alter the composition of the intestinal microbiota.
Giardia‐lamblia infection in hospitalized children with irritable bowel syndrome and dyspepsia: a prospective study
Authors: Guzyeyeva Gloria V
Published: June 2021

AIM

To evaluate the prevalence of Giardia lamblia infection in patients with irritable bowel syndrome (IBS) and dyspepsia.

METHODS

We determined G. lamblia antigens in 102 children using enzyme‐linked immunosorbent assays (ELISA). We studied the sonographic appearance of the duodenum and colon in giardiasis compared with healthy subjects.

RESULTS

Giardial lesions of the duodenum and colon were associated with increased wall thickness (p<0.05) and were characterized by increased wall echogenicity, flattening or loss of duodenal folds and/or colonic haustration, hyperechoic floating foci (HFF) demonstrating chaotic motility, increased peri‐lesional tissue echogenicity, and altered colonic peristalsis. The prevalence of IBS defined according to Rome criteria was higher in girls (28 vs 17% in boys, p = 0.001). Colonic pain frequency was similar in both genders (14% in girls vs 12% in boys, p = 0.56), however disturbed defecation occurred significantly more often in girls than in boys (19% vs 8%, p = 0.006). Half of the subjects had also dyspeptic symptoms, lactose intolerance and steatorrea. In addition, skin lesions and alopecia were determined in 6% of the patients.

CONCLUSION

Sonographic features of giardial lesions of the duodenum and colon were distinct from healthy subjects. IBS is common syndrome with the prevalence rates similar to those described in USA. [NIH grant, Russian Federation].
Interleukin‐6 as a Key Regulator for Early Hepatic Stellate Cell Transdifferentiation
Authors: Moore Cathy C. , Lakner Ashley M. , Gulledge Alyssa A. , Schrum Laura W.
Published: June 2021

Hepatic stellate cell (HSC) transdifferentiation is critical to the development of fibrosis. Phenotypic changes have been documented over the course of HSC activation; however, little information is known about daily genetic alterations. In this study we examined the expression profile of 21 genes from freshly isolated HSCs and across 10 days in culture that are important mediators of HSC activation and fibrosis by RealTime PCR and clustered the data using AMADA software. By examining specific gene activation/inhibition as well as the rise and fall of expression across time, we identified distinct markers for quiescence (GFAP and PPARγ), early and late activation (SMA, desmin, MMP‐13, IL‐6), and fibrosis (collagen, TGFβ, TGFβR and PDGFR). Our results indicated an early transient spike in the IL‐6 signaling pathway. Therefore, we chemically blocked this pathway using JAK‐2 (AG490) and p38MAPK (SB203580) inhibitors to determine if genotypic alterations correlated to changes in activation as assessed morphologically. Our results demonstrated that inhibition of JAK‐2, but not p38MAPK, impeded the progression of early HSC activation. Overall, genetic and morphological observations showed that IL‐6 signaling may be a key factor for initiation of HSC transdifferentiation. Supported by NIH Grant AA14891.
Role of histone acetylation in developmental regulation of rat GLUT5 in vivo
Authors: Douard Veronique , Mochizuki Kazuki , Goda Toshi , Ferraris Ronaldo P
Published: June 2021

In suckling mammals, expression of the intestinal fructose (F) transporter GLUT5 is low and cannot be stimulated by dietary F. When rats wean at ~14 d of age, GLUT5 transcription can be enhanced by F, indicating that regulation of GLUT5 is age‐sensitive. Histone H3 acetylation has previously been associated with transcriptional activation of GLUT5 in Caco2 cells. We perfused intestines of 10 and 20 d old rats either F or glucose (G), then determined levels of acetylation by ChIP assays using antibodies against acetylated histones H3 and H4. We used primer pairs targeting promoter regions 0 to ‐5100 bp upstream of the start site. GLUT5 expression and fructose uptake were similar among G‐ and F‐perfused intestines of 10 d and of G‐perfused 20 d olds but increased in F‐perfused intestines of 20 d olds. Histone H3 acetylation was clearly enhanced by F perfusion, but only in 20 d old rats. Increased H3 acetylation by F affected the downstream (‐1200 to 0) more than the upstream (‐5100 to ‐1400 bp) promoter regions. Likewise, histone H4 acetylation occurred only in F‐perfused intestines of 20 d olds. However, levels of H4 were lower than those of H3 acetylation and were evenly distributed between regions ‐5100 and 0 bp. F perfusion had no effect on H3 and H4 acetylation of GLUT5 in 10 d olds. Age‐dependent, F‐induced activation of GLUT5 transcription in vivo may be linked to age‐dependent factors modulating histone acetylation. (NSF722365)
CARBONYLATION CONTRIBUTES TO SERCA2 ACTIVITY LOSS DURING DIABETES
Authors: Shao ChunHong , Capek Haley L , Patel Kaushik , Nagai Ryoji , Horiuchi Seikoh , Bidasee Keshore R
Published: June 2021

The present study was undertaken to determine if carbonylation of SERCA2 contributes to the prolongation of active cardiac relaxation time during diabetes. Type 1 diabetes was induced in male Sprague‐Dawley rats using streptozotocin (STZ). After seven weeks, serum reactive carbonyl species (RCS) increased 300%. Cardiac transmitral valve flow pattern (E:A ratio) and myocyte relaxation rate were decreased by 25% and 22%. Myocyte evoked Ca2+‐transient decay time increased 3.8‐fold. Ca2+ uptake and SERCA2 Ca2+ATPase activity were reduced by 57% and 60%. SERCA2 expression decreased by 10% but phospholamban levels remained unchanged. Trypsin digestion followed by mass spectroscopic analysis revealed elevated levels of carbonyl adducts on select basic residues of SERCA2. Crosslinking pentosidine adducts were also found on SERCA2 using adduct‐specific antibodies. Incubating hSERCA2 with the methylglyoxal reduced its ability to transport Ca2+. Arg164(Y)E, Arg636(Y)E, Lys475(Y)E and Lys481(Y)E mutations to mimic adducts formed during diabetes reduced the ability of SERCA2 to transport Ca2+. Treating STZ‐diabetic rats with pyridoxamine for five weeks, starting two weeks after STZ injections reduced serum RCS levels and blunted SERCA2 activity loss without altering steady state levels of SERCA2 or phospholamban. Pyridoxamine treatment also attenuated carbonylation of SERCA2. These new data demonstrate that carbonylation of SERCA2 is functionally important and contributes to the prolongation of active cardiac relaxation time during diabetes (SUPPORTED BY ADA)
Inhibition of inflammatory gene expression in adipose tissue and development of glucose intolerance in DIO mice by a chemokine receptor 2/5 dual antagonist
Authors: Yu Xiaohong , Wang Guoming , Hamdouchi Chafiq , Kowala Mark , Zimmerman Karen , Singh Jaipal
Published: June 2021

Obesity in humans and animal models is associated with high levels of inflammatory cell infiltration in the adipose tissue. It is considered that inflammatory cell products modulate cytokine and adipokine secretion that may contribute to the pathogenesis of insulin resistance, diabetes and atherosclerosis. In the present study we show that diet induced alteration in inflammatory gene expression in the adipose is reduced by treatment with a chemokine (C‐C motif) receptor 2/5 (CCR2/CCR5) dual antagonist. Gene expression analysis of adipose tissue of C57/BL6 mice subjected to high fat diet for 6 weeks showed a significant increase in the expression of TNF a, MCP‐1, CD68, and cathepsin S. Treatment with a CCR2/CCR5 antagonist starting two weeks after the initiation of high fat diet significantly reduced (>70%, p< 0.05) the expression of these genes in the adipose. Measurement of glucose tolerance at the end of six weeks showed a dose‐dependent improvement in glucose disposal in the treated groups. At 30 mg/kg of antagonist dose, the glucose disposal rate became similar to that of the normal lean animals. In addition, CCR2/CCR5 antagonist produced 40 % reduction (p<0.05) in liver triglycerides without affecting inflammatory gene expression in the liver. These data show that in a preclinical model of diet induced obesity (DIO), a potent CCR2/CCR5 dual antagonist inhibited adipose inflammatory gene expression, reduced adipose fat mass and improved insulin sensitivity.
Cardiac Phenotypic Differences in Rat Models of the Metabolic Syndrome
Authors: Ohanyan Vahagn , Pung Yuh Fen , Hafemeister Jennifer L , Hodnichak Cheryl , Kolz Christopher , Chilian William M
Published: June 2021

In humans, the metabolic syndrome denotes a constellation of risk factors including obesity, insulin resistance, hypertension, dyslipidemia, and pro‐inflammatory and pro‐thrombotic states. Patients with some or all of these major symptoms have a high risk of developing heart failure. To elucidate if animal models of the metabolic syndrome also demonstrate early signs of ventricular pathology, we studied age‐matched JCR, Zucker Obese Fatty (ZOF) (both strains used as models for the metabolic syndrome), and Lean (LN) rats. Measurements of cardiac function and left ventricular parameters (echocardiography) were made when the animals were lightly anesthetized with sevoflurane (refer to table mean±SD [Dias‐ID: diastolic internal diameter, mm; RWT: relative wall thickness; LV Mass, mg; CI: Cardiac Index, ml/min/kg, %EJ: ejection fraction).

These results support the idea that either model shows varying degrees of ventricular pathology as evidenced by the reduced cardiac index and increased mass (both strains), but the extent of dysfunction appears to be more severe in JCR because of ventricular dilation and reduced ejection fraction.
Tumor necrosis factor‐alpha production by peripheral blood dendritic cells in post menopausal type 2 diabetic women
Authors: Blank Sally Elliott , Johnson Emily Carolyn , Weeks Debbie , Wysham Carol H.
Published: June 2021

The objective was to examine if tumor necrosis factor‐alpha (TNF‐α) production by subsets of blood dendritic cells (DC), plasmacytoid (pDC) and myeloid (mDC), is altered in post‐menopausal women with type 2 diabetes (T2D).Women (n=21) were recruited from a local sample enrolled in the Action to Control Cardiovascular Risk in Diabetes (ACCORD) clinical trial. Plasma glucose and TNF‐α were determined by amperometry and ELISA, respectively. Intracellular TNF‐α was analyzed by flow cytometry. T2D had greater (p<0.05) BMI, plasma glucose and TNF‐α vs healthy women (n=21). Poor glycemic control (HA1c >7%) was present in 67% of T2D. Age and glycemic control (GLYC) effects, examined by ANOVA, indicated that plasma TNF‐α was greater in T2D with poor GLYC, independent of age (p=0.004). An interaction existed between age and GLYC (p=0.041) with a trend (p=0.07) for lower mDC TNF‐α in older women (age > 60 yr) with poor GLYC. T2D with plasma TNF‐α >15pg/mL had fewer stimulated mDCs (%) expressing TNF‐α vs healthy women (p=0.045). The findings support the hypothesis that TNF‐α production by DCs is modified by glycemic control in women with T2D. Support by McLeod Endowment Fund, WSU College of Pharmacy.
Glucocorticoids increase basal lipolysis, but not epinephrine stimulated lipolysis
Authors: Campbell Jonathan , Hawke Thomas J , Riddell Michael C
Published: June 2021

Glucocorticoids (GCs) are associated with central obesity, yet have been classically defined as lipolytic. We have recently shown that exercise training decreases fat mass while increasing GC action in adipocytes, perhaps by increasing adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL). From this, we hypothesize that the lipolytic effects of GCs are mediated through changes in these adipose enzymes. To test this, we treated 3T3‐L1 adipocytes with GCs for 48 hours, and then allowed the cells to undergo basal or epinephrine stimulated lipolysis. 48 hours of GCs modestly increased lipolytic rates (1.6 fold vs con, p<0.05). However, the basal lipolysis following GC treatment was substantially increased (3.5 fold vs. con, p<0.01). The expression of ATGL was increased 3.5 fold over control (p<0.01), whereas HSL expression was increased only 1.5 fold (p<0.05). The phosphorylation of HSL at Ser563 and Ser660, sites that increase HSL activity, were also increased significantly (4.5‐and 2.2‐fold vs con, respectively; p<0.01). GC treatment only modestly increased epinephrine stimulated lipolysis (1.5 fold vs con, p<0.05). The phosphorylation of HSL at Ser563 and Ser565 by epinephrine was not affected by GC treatment. Thus, it appears GCs influence lipolysis mainly through alterations in basal lipolysis predominantly by altering ATGL expression and HSL activity. Supported by NSERC and CIHR.
PDK‐1 acts upstream of PKC‐delta to mediate augmented endothelin‐1 vasoconstriction in mesenteric arteries from rats exposed to intermittent hypoxia
Authors: Webster Bradley R , Kanagy Nancy L
Published: June 2021

Mimicking sleep apnea by exposing rats to eucapnic intermittent hypoxia (E‐IH) (3 min cycle of 5% O2, 5% CO2/air flush 7 hr/day 2 wks) increases blood pressure and endothelin‐1 (ET‐1) levels compared to controls (air/air cycles 7hr/day 2 wks). We have shown that augmented ET‐1 constriction in denuded mesenteric arteries (MA) is through PKCd in E‐IH but not Sham MA. Classical PKC isoforms play a minimal role. In intact MA, rottlerin (3 μM), a PKCd inhibitor, attenuated ET‐1 constriction in E‐IH (79±3.6% w/o rottlerin 40.3 ±8.9% w/*) but not Sham MA (69.3 ± 4.3% w/o rottlerin 65.5 ± 4.8% w/). PKC is primed for interaction with ET‐1 generated DAG by PDK‐1. We hypothesized that i[NLK1]nhibiting PDK‐1 with OSU‐03012 (10 μM) would be similar to PKCd inhibition. OSU‐03012 attenuated constriction in E‐IH (79.4 ±4.4% w/o OSU 55.8 ±8.1% w/*) but not in Sham MA (72.3 ± 7.1% w/o OSU 71.8 ± 3.3% w/ OSU). Western blots showed OSU‐03012 decreased the total PKCd expression in E‐IH but not Sham MA suggesting PDK‐1 stabilizes PKCd. PDK‐1 can be activated by PI‐3K. PI‐3K inhibition with LY294002 (10 μM) did not affect constriction in E‐IH vessels (75.8±6.6% w/o LY 78.3 ±3.3% w/ LY) and augmented constriction in Sham MA (69.2±5.6% w/o LY 85.4 ±5.5% w/, p = .09). Therefore augmented constriction in E‐IH MA is not through PI‐3K activation of PDK‐1 . In summary, PDK‐1 appears to increase PKCδ stability resulting in augmented E‐IH MA vasoconstriction to ET‐1. (* = p < .05)
β‐adrenergic stimulation of K secretion in guinea pig distal colon through activation of both Cl and K conductances
Authors: He Quanhua , Zhang Jin , Halm Susan , Halm Dan
Published: June 2021

K secretion was measured as whole‐cell currents in isolated colonic crypts as well as short‐circuit current (Isc) and transepithelial conductance (Gt) in guinea pig distal colonic mucosa. Colonic crypts isolated from basement membrane were sealed with patch pipets and standard whole cell obtained. Currents recorded at K reversal potential were defined as predominantly Cl currents (ICl) and those at Cl reversal potential were K currents (IK). Epinephrine (epi) [10μM] stimulation resulted in a 1.4‐fold increase in IK that was maximal within 2min; a 1.8‐fold increase in ICl occurred that reached maximal levels within 5min. Epi stimulated negative Isc consistent with K secretion that was near maximum by 10min; Gt increased in parallel with Isc activation. K secretory Isc was largely inhibited by addition of glibenclamide (glib) to either mucosal or serosal bathing solution with an EC50 of ~200μM. Addition of glib to both sides increased sensitivity, EC50 ~40μM. Glib [100μM] eliminated epi activated IK and ICl in crypts within 2min; ICl decreased further over the succeeding 10min indicating that basal ICl also was partially glib‐sensitive. Mucosal expression was detected by RT‐PCR for glib‐sensitive K channel proteins Kir6.1 and Kir6.2 and accessory subunits SUR1 and SUR2. Thus, K secretion likely involves glib‐sensitive K and Cl channels present in apical and basolateral membranes of colonic crypts. [NIH DK65845]
Tamoxifen inhibits TRPV6 activity via estrogen receptor independent pathways in TRPV6 transfected MCF‐7 cells
Authors: Bolanz Katrin Annika , Kovacs Gergely Gyula , Landowski Christopher Paul , Hediger Matthias A
Published: June 2021

TRPV6 expression has been shown to be upregulated in breast cancer tissue compared to normal mammary gland tissue. In several cell lines, it was shown that knockdown of TRPV6 inhibits cell growth. Tamoxifen is a selective estrogen receptor modulator widely used in breast cancer therapy. Previously, we showed that tamoxifen down regulates TRPV6 mRNA expression and that it also inhibits calcium uptake in TRPV6‐transfected oocytes. In the present study we examined the effect of tamoxifen on TRPV6 function in MCF‐7 breast cancer cells transiently transfected with TRPV6, tagged with EYFP. TRPV6 function was measured with fluorescence microscopy using Fura‐2. The influx of calcium, barium and manganese was significantly larger in cells transfected with this construct, compared to non‐transfected cells. In transfected cells, 10 µM tamoxifen induced a decrease in intracellular calcium concentration in transfected cells down to the basal calcium level of non‐transfected cells, following a large, transient increase. In nominally calcium free buffer tamoxifen induced a smaller transient increase. The transport rate of calcium and barium but not manganese was decreased 50‐70% by tamoxifen. This inhibitory effect was not mediated by estrogen receptors as the antiestrogen ICI 182,720 had no inhibitory effect. Inhibition of PI3K with 25 µM LY294002 had no effect on TRPV6 activity. These findings give novel insights into how tamoxifen might be effective in estrogen receptor negative breast cancer patients. Our data suggest that TRPV6 is involved in the therapeutic mechanism of tamoxifen during breast cancer treatment. This study was supported by the Novartis Stiftung and the Bernische Krebsliga (PI's Hediger and Landowski).
Anion secretion evoked by ATP in rabbit cornea and its suppression in dry eye model
Authors: Yu Haijie , Xiong Cuiju , Chen Silinag , Ma Lijuan , Ruan Yechun , Wang Zhichong , Zhou Wenliang
Published: June 2021

To explore the regulation of anion secretion across rabbit corneal epithelia by extracellular ATP and to investigate the mechanism of benzalkonium chloride (BAC) induced dry eye model involved in this secretion, short circuit current and patch clamp were used. Changes in cytosolic Ca2+ concentration were measured using Fluo3. The cornea responded to apical application of ATP in short circuit current (Isc). Ion substitution experiments showed the rise in Isc due to Cl‐ and HCO3‐ secretion. Pretreatment with Cl‐ channel blockers, such as DIDS, NPPB and DPC attenuated the response. Moreover, the Isc was inhibited by U73122 and BAPTA/AM, indicating the requirement of PLC and intracellular calcium (Ca2+i) in the anion secretion. In whole‐cell patch clamp study, ATP stimulated a Ca2+‐activate Cl‐ channel and this current showed an outward rectifying I‐V relationship. In BAC‐pretreated epithelial cell, the whole‐cell current was suppressed. Dynamic changes of Ca2+i demonstrated the elevations of Ca2+i was smaller in BAC‐treated epithelia compared to control cells when they were challenged by ATP and Thapsigagin. These findings suggested ATP could induce anion secretion, and this secretion was inhibited in BAC‐evoked rabbit dry eye model. Abnormality of intracellular Ca2+ and Ca2+ activated Cl‐ channel may take charge of inhibitory anion secretion in the rabbit corneal epithelial cells of dry eye model.
Comparison of personal formaldehyde levels in the anatomy laboratories of five physical therapy education programs
Authors: Cope Janet M. , Sanders Erin , Holt Stewart M , Pappas Kathy , Thomas K. Jackson , Kernick Edward , Corriher Laura , Templeton Davis
Published: June 2021

Formaldehyde (FA) has been classified by The International Agency for Research on Cancer as a human carcinogen (Levine et al, 1984; IEQ, 2006; Kriebel, 2001) and as a probable carcinogen by the Environmental Protection Agency (1999). The purpose of this research was to compare personal FA exposure and sensory irritation levels between subjects in the human anatomy laboratories at five Physical Therapy (PT) programs in the United States. There was no significant correlation between FA levels and irritation levels. There was not a significant difference in FA exposure between students and instructors but there was a significant difference (p=0.015) in irritation levels reported by students and instructors, with instructors reporting lower irritation levels. There was a significant difference (p=0.001) in irritation levels between institutions but no significant difference in FA exposure levels between institutions. At one institution the FA levels in donors that were reperfused with Infutrace were significantly lower (p=0.002) than those of donors that were non‐reperfused. Future studies that compare teaching strategies, dissection techniques, and ventilation systems should be conducted to help determine the healthiest combination of methods (chemicals, ventilation systems, teaching strategies, personal protective equipment) to reduce FA and irritation levels among students and instructors.
The limitation of plasma volume correction for the serum protein concentration analysis in an acute exercise
Authors: JANG SEONWOONG , KIM DONGHEE , LEE HAYAN , BAEK KYOUNGYEOP , YANG MEEHYUN , KIM HYEOK , KWON SOYOUNG , KIM DAEYEOL , JUNG HYEMIN , CHOI CHEOLSEUNG
Published: June 2021

An acute exercise concomitant with transient plasma shifts caused by the modified colloid osmotic pressure gradient contributes to the plasma volume correction(PVC) demands. However, relations between physiological effects of an exercise and the physical characteristics of permeating molecules or endothelial barriers raise doubts about quantitative benefits of PVC. The present study was conducted to determine the limitation of widely applied PVC on measuring concentration of serum protein in an acute exercise. Nine college men(age: 25.4±0.5, BMI: 22.6±1.9) were assigned to non‐exercise group(NG) on the first day and to exercise group(EG)(45 min., 65‐75 %HRmax, running) on the next day. Blood samples were drawn for the serum protein electrophoresis analysis(t‐test, pearson correlation) twice(pre‐event, post‐event). Albumin(p<.05), α1(p<.01), α2(p<.01), β(p<.05), and λ(p<.05) fraction concentration increased after the exercise. Negative correlations between the percentage of change in albumin and that of other fractions concentration in NG(α1(r=‐.755, p<.05), α2(r=‐.947, p<.01), β(r=‐.954, p<.01), λ(r=‐.928, p<.01)) decreased by wide gaps in EG(α1(r=.377), α2(r=‐.210), β(r=‐.349), γ(r=‐.289)). These correlations moved from the positive direction to the negative according to the increasing size of serum proteins in both groups except for γfraction. The results suggest that not only relatively small‐sized albumin but large‐sized serum proteins permeate endothelial barriers with conformational strain in proportion to the size. Therefore, the application of current PVC on a study measuring serum protein concentration in an acute exercise may underestimate the accurate concentration in situ.
Lactobacillus acidophilus supplementation for 7 days does not reduce acute aspirin‐induced gastrointestinal permeability
Authors: Lambert G. Patrick , Bedard Nicholas , Holte Gina , Schmidt Angela , Lanspa Stephen
Published: June 2021

The purpose of this study was to determine the effect of Lactobacillus acidophilus (L. acidophilus; LA) on aspirin‐induced gastrointestinal (GI) permeability. Fourteen subjects (8 male; 6 female; age = 21 ± 0.4 yrs) completed three trials: 1) LA placebo/aspirin, 2) LA placebo/aspirin placebo, and 3) LA/aspirin. Subjects ingested either LA (10 billion colony forming units per day) or placebo for seven days. On the evening of the sixth day and morning of the seventh day they ingested aspirin (1300 mg) or aspirin placebo. Subjects then drank a solution (150 ml) containing 5 g sucrose, 5 g lactulose, and 2 g rhamnose and urine was collected for 5 hours. Gastroduodenal permeability was determined from urinary sucrose excretion and small intestinal permeability was determined from the lactulose‐to‐rhamnose urinary excretion ratio. Gastroduodenal permeability was significantly (p < 0.017) greater in the LA placebo/aspirin and LA/aspirin trials compared to the LA placebo/aspirin placebo trial. Small intestinal permeability was also significantly (p < 0.017) greater in the LA placebo/aspirin and LA/aspirin trials compared to the LA placebo/aspirin placebo trial. These results indicate that aspirin ingestion increases GI permeability and 7 days of LA does not reduce this effect. Supported by the Gatorade Sports Science Institute. Danisco USA, Inc. provided the L. acidophilus.
Acute ethanol exposure aggravates cockroach‐allergen (CRA) induced asthmatic responses
Authors: Bouchard Jacqueline Claire , Kim Jiyoun , Vaickus Louis , Remick Daniel
Published: June 2021

This study investigates the effects of acute ethanol exposure on CRA induced murine asthma. Outbred mice were immunized intratracheally with CRA and challenged 2 times (Day 14 and 21). On Day 21 the Ethanol and Water groups were given 300µl 32% ethanol or water by oral gavage 30 minutes prior to second challenge (SC). Blood ethanol levels at the time of SC were 0.1%. Five hours post‐SC blood was collected for IgE measurements and lungs lavaged for cell counts and chemokine/cytokine measurements. Alcohol exacerbated the asthmatic inflammatory response by a) increasing bronchoalveolar lavage (BAL) chemokines Eotaxin‐2 and RANTES with corresponding increases in eosinophil and lymphocyte BAL infiltrates, b) increased total and CRA‐specific IgE plasma levels, and c) skewed towards a TH2 inflammatory response by decreasing BAL IFNγ while increasing BAL IL‐4, IL‐5, and IL‐13. Supported by 5R01ES013538‐04.
Concentration of transforming growth factor‐beta in an infant formula from five countries and human milk from Mexico and the US
Authors: Jouni Zeina E , Waworuntu R. , Vangari G. , Slightom R. , Rosales R. , Ferguson P. , Rai G. , Ruiz‐Palacio G. , Guerrero M. , Morrow A. , McMahon R.
Published: June 2021

Transforming growth factor‐beta (TGF‐beta) is found in human milk and is thought to be a key growth factor for the development of the immune system and oral tolerance in neonates. In this study, the concentration of total TGF‐beta (isoforms beta‐1 and beta‐2) was determined in human milk obtained from mothers in the US and Mexico, and compared to that found in an infant formula (Enfamil LIPIL powder) marketed in 5 countries. Human milk samples (n=260) were collected at specific times over the first 26 weeks of lactation from three cohorts of 20 healthy mothers who delivered term babies. Infant formula powder samples (n=25) representing multiple manufacturing sites were obtained from the US, Mexico, Thailand, China, and Philippines. TGF‐beta isoforms were quantified by a specific ELISA method validated for each type of sample. When pooled across the cohorts and collection times, human milk exhibited a geometric mean of 2,700 pg/ml and a 95% confidence interval of 550 ‐ 13,000 pg/ml. The mean concentration of TGF‐beta in the reconstituted infant formula was 4,900 pg/ml with a range of 2,800 ‐ 9,900 pg/ml. These data demonstrate that TGF‐beta is present in infant formula at a level within the range found in human milk.
Reflecting on two distinct leadership roles during the pandemic
Authors: Kroning Maureen
Published: June 2021

For more than a year, the COVID-19 pandemic has caused unprecedented stress, anxiety, sadness, and fear, but it has also allowed people to reflect, learn, self-improve, and change. In this article, the author demonstrates how reflective practice enabled her to improve in two distinctly different leadership roles.
Posttraumatic stress disorder in nurses caring for patients with COVID-19
Authors: Hill Richard
Published: June 2021

Little is known about the psychological impact of trauma from pandemics such as the COVID-19 pandemic. This article explores a descriptive study on the impact of COVID-19 and the prevalence of posttraumatic stress disorder among RNs caring for patients with COVID-19.
The emerging role of robotics in upper and lower urinary tract reconstruction
Authors: Zhao Calvin C. , Shakir Nabeel A. , Zhao Lee C.
Published: June 2021

Purpose of review

Developments in robotic reconstructive urology have introduced novel treatments for complex upper and lower urinary tract disease. Short-term and mid-term data demonstrates excellent outcomes and minimal morbidity, suggesting the advanced instrumentation and visualization of robotics represent a new treatment paradigm in patients that are historically difficult to treat. Here we review recent developments in the robotically assisted surgical management of urethral and ureteral strictures.

Recent findings

The minimally invasive approach, enhanced precision and reach, and near-infrared fluorescence imaging capabilities of robotic platforms have proven to be valuable additions in reconstructive urology where perfusion is often compromised, or anatomy is distorted. These benefits are leveraged heavily in recent descriptions of robotic-assisted posterior urethroplasty and ureteroplasty. Short-term to mid-term follow-up data for these procedures show excellent patency rates with low morbidity and complication rates when compared with open approaches. Long-term data for these procedures are not yet available.

The role of robotics in reconstructive urology is being actively investigated. Initial findings demonstrate excellent results with low morbidity in the treatment of upper and lower urinary tract disease. Long-term data will ultimately determine the role of robotics in the reconstructive armamentarium.
Triphasic Waves: Historical Overview of an Unresolved Mystery
Authors: Fernández-Torre José L. , Kaplan Peter W.
Published: June 2021

Triphasic waves are a fascinating and mysterious EEG feature. We now have to accept that, at times, epileptiform discharges may have a blunted “triphasic morphology,” and that there may be great difficulty in distinguishing between these often similar forms. The aim of this review was to describe the evolution in our understanding of triphasic waves that has occurred regarding the pathophysiology of triphasic waves, their most frequent causes, and the diagnostic difficulties involved in interpretation and differentiation from nonconvulsive status epilepticus.
Assessing ethnic business sustainability in transgenerational succession
Authors: Xavier Jesrina Ann , Feranita Feranita , Jambulingam Manimekalai , Gorchan Singh Manmeet Kaur
Published: June 2021

Purpose

This paper aims to examine the impact of changes in human capital development and evolution of tacit knowledge following transgenerational succession in ethnic companies. The paper contributes to the understanding of transferring tacit knowledge across generations in ensuring ethnic business sustainability.

Design/methodology/approach

In answering the how question, this paper tracked the changes and their impact in the process over time, using the multiple-case study method. A total of six interviews were conducted with three Indian-owned companies in the jewellery industry in Malaysia, with each interview lasting between 45 and 60 min. Secondary data were collected to supplement the primary data for analysis. Data triangulation method was applied to strengthen the design of this study.

Findings

The results indicate that changes in human capital development and tacit knowledge have enabled ethnically Indian-owned jewellery-based companies to alter their products to respond to demands of modern society whilst sustaining and commodifying the ethnic identity of their businesses. The findings also highlight that proper succession planning by ageing entrepreneurs may promote sustainability of these ethnic enterprises.

Originality/value

Despite the growing attention on ethnic and migrant entrepreneurship, less is known about the impact of the changes through transgenerational succession over time in ethnic businesses, especially when such changes involve human capital as the key players. This study is important in addressing the gap, in identifying human capital development and tacit knowledge among the critical ethnic resources contributing to ethnic business sustainability. Using a conceptual framework, this paper sheds some light on how ethnic businesses are sustained through transgenerational succession.
Supply chain agility responding to unprecedented changes: empirical evidence from the UK food supply chain during COVID-19 crisis
Authors: Do Quynh Nhu , Mishra Nishikant , Wulandhari Nur Baiti Ingga , Ramudhin Amar , Sivarajah Uthayasankar , Milligan Gavin
Published: June 2021

Purpose

The COVID-19 outbreak has imposed extensive shocks embracing all stages of the food supply chain (FSC). Although the magnitude is still unfolding, the FSC responds with remarkable speed, to mitigate the disruptive consequences and sustain operations. This paper aims to investigate how operationalising supply chain agility (SCA) practices has occurred amid the COVID-19 crisis and expectations for how those practices could transform the supply chain in the post-COVID-19 era.

Design/methodology/approach

Following an exploratory case-based design, this paper examines the various agile responses that three supply chains (meat, fresh vegetables and bread) adopted and elaborate using the dynamic capability (DC) theoretical lens.

Findings

First, the findings demonstrate how, in the wake of the COVID-19 pandemic, each affected case pursued various agile responses through sensing and seizing capabilities. Sensing includes identifying and assessing the relevant opportunities and threats associated with the specific supply chain context. Seizing involves acquiring, combining and modifying the tangible and intangible resources at the firm and supply chain levels. Second, supply chain transformation is likely if firms and their supply chain develop the sustaining capability to ensure that the desirable changes outlast the crisis.

Practical implications

This study provides an actionable guide for practitioners to develop agile responses to systemic changes in times of crisis and to sustain favourable changes so as to enable their outlasting of the crisis.

Originality/value

This study provides a novel and unique perspective on the role of SCA in crisis – in this case, the pandemic. This paper synthesises the empirical stories of the agile responses in the FSC and elaborates on the DC framework, to identify theoretical and practical implications. This paper establishes the sustaining capability as the missing DC capability for enabling transformation in the post-COVID-19 era.
Implementing fast track surgery in hip and knee arthroplasty using the lean Six Sigma methodology
Authors: Latessa Imma , Fiorillo Antonella , Picone Ilaria , Balato Giovanni , Trunfio Teresa Angela , Scala Arianna , Triassi Maria
Published: June 2021

Purpose

One of the biggest challenges in the health sector is that of costs compared to economic resources and the quality of services. Hospitals register a progressive increase in expenditure due to the aging of the population. In fact, hip and knee arthroplasty surgery are mainly due to primary osteoarthritis that affects the elderly population. This study was carried out with the aim of analysing the introduction of the fast track surgery protocol, through the lean Six Sigma, on patients undergoing knee and hip prosthetic replacement surgery. The goal was to improve the arthroplasty surgery process by reducing the average length of stay (LOA) and hospital costs

Design/methodology/approach

Lean Six Sigma was applied to evaluate the arthroplasty surgery process through the DMAIC cycle (define, measure, analyse, improve and control) and the lean tools (value stream map), adopted to analyse the new protocol and improve process performance. The dataset consisted of two samples of patients: 54 patients before the introduction of the protocol and 111 patients after the improvement. Clinical and demographic variables were collected for each patient (gender, age, allergies, diabetes, cardiovascular diseases and American Society of Anaesthesiologists (ASA) score).

Findings

The results showed a 12.70% statistically significant decrease in LOS from an overall average of 8.72 to 7.61 days. Women patients without allergies, with a low ASA score not suffering from diabetes and cardiovascular disease showed a significant a reduction in hospital days with the implementation of the FTS protocol. Only the age variable was not statistically significant.

Originality/value

The introduction of the FTS in the orthopaedic field, analysed through the LSS, demonstrated to reduce LOS and, consequently, costs. For each individual patient, there was an economic saving of € 445.85. Since our study takes into consideration a dataset of 111 patients post-FTS, the overall economic saving brought by this study amounts to €49,489.35.