CAL Group Research Articles

The mechanism of intravenous immunoglobulin (IVIG) in vascular endothelial injury in kawasaki disease based on neutrophil extracellular traps

This study explores the role of Neutrophil extracellular traps (NETs) in kawasaki disease (KD)-induced vascular inflammatory injury and the protective effect and mechanism of IVIG on vascular endothelial damage. A total of 37 children diagnosed with KD and admitted to Dongguan maternal and Child Health Care Hospital between March 2020 and June 2022 were included in the study. The children were divided into different groups based on their treatment and the presence or absence of coronary artery damage: IVIG treatment group (KDIVIG group), subgroup with coronary artery damage (KDCAL group), and subgroup without coronary artery damage (KDNCAL group), and a Control group consisting of 9 children who underwent surgical treatment. Flow cytometry was used to detect the proportion of neutrophils and the number of NETs in peripheral blood. It was found that the proportion of neutrophils in the peripheral blood of the acute KD group significantly increased with the presence of NETs. RT-PCR and ELISA detection showed that the levels of inflammatory factors TNF-α, IL-6 and CitH3 were abnormally elevated in this acute KD group, and the CAL group exhibited higher proportions of neutrophils and NETs-related markers compared to the NCAL group, while the IVIG group had significantly decreased proportions of neutrophils. PMA culture of neutrophils induced an increase expression of NETs marker protein, the content of NETs cfDNA increased. NETs culture could promote the secretion of TNF-α, whereas IVIG cultured cells inhibited the secretion of TNF-α. Finally, HCAEC cells were cultured with different levels of TNF-α, and the function of HCAEC cells was assessed using CCK8, scratch assay and flow cytometry. The high expression of TNF-α in the NETs group inhibited the proliferation and migration of HUVEC cells and enhanced their apoptosis. In contrast, the IVIG culture group exhibited similar effects to the TNF-α monoclonal antibody, as it inhibited HUVEC cell apoptosis and improved their viability by reducing TNF-α expression. Total protein was extracted from the cells using nano-magnetic beads, and RT-PCR and western blot detection indicated that the increase of TNF-α expression could increase the phosphorylation of NF-κB and and the expression of MMP-9. However, when TNF-α was inhibited by IVIG and TNF-α monoclonal antibody culture, the activity of NF-κB/MMP-9 athway was decreased. Therefore, IVIG may inhibit the production of NETs in KD children, thereby reducing TNF-α/NF-NF-κB/MMP-9 mediated inflammatory response process and protecting the function of vascular endothelial cells.

Calcitriol promotes M2 polarization of tumor-associated macrophages in 4T1 mouse mammary gland cancer via the induction of proinflammatory cytokines

Our research found that vitamin D3 (VD3) treatment increased lung metastasis in mice with 4T1 murine breast cancer (BC). This study aims to investigate the impact of VD3 on the activation of tumor-associated macrophages (TAMs) in BC. Mice bearing 4T1, E0771, 67NR BC cells, and healthy mice, were fed diets with varying VD3 contents (100—deficient, 1000—normal, and 5000 IU/kg—elevated). Some mice in the 1000 and 100 IU/kg groups received calcitriol. We studied bone metastasis and characterized TAMs and bone marrow-derived macrophages (BMDMs). 4T1 cells had higher bone metastasis potential in the 5000 IU/kg and calcitriol groups. In the same mice, an elevated tumor osteopontin level and M2 polarization of TAMs (MHCIIlow CD44high phenotype) were observed. Gene expression analysis confirmed M2 polarization of 4T1 (but not 67NR) TAMs and BMDMs, particularly in the 100 IU + cal group (increased Mrc1, Il23, and Il6). This polarization was likely due to COX-2/PGE2 induction in 4T1 calcitriol-treated cells, leading to increased proinflammatory cytokines like IL-6 and IL-23. Future studies will explore COX-2/PGE2 as a primary mediator of calcitriol-stimulated inflammation in the BC microenvironment, especially relevant for BC patients with VD3 deficiency and supplementation.

Bioactive potential of Bio-C Temp demonstrated by systemic mineralization markers and immunoexpression of bone proteins in the rat connective tissue

Intracanal medications are used in endodontic treatment due to their antibacterial activity and ability to induce the periapical repair. Among the intracanal medications, the Calen (CAL; SS. White, Brazil) is a calcium hydroxide-based medication that provides an alkaline pH and releases calcium, exerting an antimicrobial activity. Bio-C Temp (BIO; Angelus, Brazil), a ready-to-use bioceramic intracanal medication, was designed to stimulate the mineralized tissues formation. Here, we investigated the bioactive potential of BIO in comparison to the CAL in the rat subcutaneous. Polyethylene tubes filled with medications, and empty tubes (control group, CG) were implanted in the subcutaneous tissue of rats. After 7, 15, 30 and 60 days, the blood was collected for calcium (Ca+2) and alkaline phosphatase (ALP) measurement, and the capsules around the implants were processed for morphological analyses. The data were submitted to two-way ANOVA and Tukey test (p < 0.05). At 7, 15 and 30 days, the ALP level was grater in BIO and CAL than in CG (p < 0.0001). At 7 and 15 days, greater Ca+2 level was seen in the serum of CAL samples. From 7 to 60 days, an increase in the number of fibroblasts, osteocalcin- and osteopontin-immunolabelled cells was observed in BIO and CAL groups (p < 0.0001). In all periods, BIO and CAL specimens showed von Kossa-positive structures. Moreover, ultrastructural analysis revealed globules of mineralization in the capsules around the BIO and CAL specimens. Thus Bio-C Temp caused an increase in the ALP, osteocalcin and osteopontin, which may have allowed the formation of calcite, suggesting bioactive potential.Graphical

Association between FGA gene polymorphisms and coronary artery lesion in Kawasaki disease.

To investigate the correlation between FGA gene polymorphisms and coronary artery lesion in Kawasaki disease. Two hundred and thirty four children with Kawasaki disease (KD group), 200 healthy children (normal group) and 208 children with non-KD fever (fever group) were enrolled. General clinical indicators, the concentration of serum MMPs, TIMP-1, FG-α,fibrinogen level, molecular function (FMPV/ODmax) and FGA Thr312Ala polymorphism were detected individually by testing peripheral venous blood after fasting in the morning. There was no significant difference in average age among the three groups, which were 3.03 ± 1.22 years, 3.17 ± 1.30 years, and 3.21 ± 1.31 years, respectively. Compared with those in the fever group, the levels of white blood cell count (WBC), platelet count (PLT), procalcitonin (PCT), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and fibrinogen (Fg) levels were significantly increased in the KD group. Red blood cell count (RBC) and hemoglobin (Hb) levels were significantly decreased (p < 0.05).The concentration of serum MMPs, TIMP-1, and FG-α in the KD and fever groups were significantly higher than those in the normal group (p < 0.05). The concentration of MMP-2, MMP-3, MMP-9, MMP-13, TIMP-1, and FG-α in the KD group were significantly higher than those in the fever group (p < 0.05).The KD group was divided into two subgroups,55 patients with combined CAL and 179 patients without combined CAL. The plasma fibrinogen concentration in the combined CAL group was significantly higher than that in the non-combined CAL and normal groups (p < 0.01). There was no statistically significant difference in FMPV/ODmax among the three groups (p > 0.05). Compared with normal group, the FGA GG, GA, and AA genotype and G, A allele frequency of the FGA gene polymorphism in the KD group showed no significant difference (p > 0.05). In the KD group, the most common type in children with CAL was GA, while the most common type in children without CAL was GG. MMPs and FG-α were significantly upregulated in KD patients. The proportion of FGA genotype GA in children with CAL was significantly higher than that in children without CAL, suggesting that FGA gene polymorphisms affect coronary artery lesion in children with KD.

Effects of calcium supplementation to resuscitation fluids in endurance horses: A randomized, blinded, clinical trial.

The addition of calcium to resuscitation fluids is a common practice in horses, but studies evaluating the effects of calcium supplementation are limited. In healthy horses, decreases in heart rate and changes in serum electrolyte concentrations have been reported. Calcium gluconate administration at a rate of 0.4 mg/kg/min to eliminated endurance horses with metabolic problems will affect heart rate, gastrointestinal sounds, and serum electrolyte concentrations. Endurance horses eliminated from the Tevis Cup 100-mile (160 km) endurance ride for metabolic problems and requiring IV fluid therapy were eligible. Sixteen horses were randomly assigned to receive 0.4 mg/kg/min of calcium (23% calcium gluconate solution) over 1 hour diluted in 10 L of a non-calcium containing isotonic crystalloid (CAL group) or 10 L of a non-calcium containing isotonic crystalloid (CON group). Staff members administering the fluids were blinded to treatment group. Blood samples were collected and physical examinations performed before and after treatment. Heart rates were recorded every 15 min during fluid administration. Data were compared using 2-way analysis of variance (ANOVA) with repeated measures for continuous variables and Fisher's exact test for categorical variables. Calcium was associated with lower heart rates 45 min after starting the infusion (P = .002). Gastrointestinal sounds were less likely to improve in the calcium group compared with the control group (P = .005). An increase in plasma phosphorus concentration (P = .03) was associated with calcium administration. Intravenous calcium supplementation to endurance horses eliminated from competition after development of metabolic problems may decrease heart rate but impairs improvement in gastrointestinal sounds.

Modulation of Fibroblast Activity via Vitamin D3 Is Dependent on Tumor Type-Studies on Mouse Mammary Gland Cancer.

Simple SummaryThis study, which was conducted in healthy mice and mice bearing three mouse mammary gland cancers—4T1, 67NR, and E0771—showed that the divergent effects of vitamin D3 supplementation (5000 IU) or deficiency (100 IU of vitamin D3) observed in healthy mice led to the formation of various body microenvironments depending on the mouse strain. Developing tumors themselves modified the microenvironments by producing higher concentrations of osteopontin, SDF-1 (4T1), TGF-β (4T1 and E0771), CCL2, VEGF, FGF23 (E0771), and IL-6 (67NR), which influences the response to vitamin D3 supplementation/deficiency and calcitriol administration and leads to enhanced/decreased activation of lung fibroblasts and modulation of tumor tissue blood flow.Vitamin D3 and its analogs are known to modulate the activity of fibroblasts under various disease conditions. However, their impact on cancer-associated fibroblasts (CAFs) is yet to be fully investigated. The aim of this study was to characterize CAFs and normal fibroblasts (NFs) from the lung of mice bearing 4T1, 67NR, and E0771 cancers and healthy mice fed vitamin-D3-normal (1000 IU), -deficient (100 IU), and -supplemented (5000 IU) diets. The groups receiving control (1000 IU) and deficient diets (100 IU) were gavaged with calcitriol (+cal). In the 4T1-bearing mice from the 100 IU+cal group, increased NFs activation (increased α-smooth muscle actin, podoplanin, and tenascin C (TNC)) with a decreased blood flow in the tumor was observed, whereas the opposite effect was observed in the 5000 IU and 100 IU groups. CAFs from the 5000 IU group of E0771-bearing mice were activated with increased expression of podoplanin, platelet-derived growth factor receptor β, and TNC. In the 100 IU+cal group of E0771-bearing mice, a decreased blood flow was recorded with decreased expression of fibroblast growth factor 23 (FGF23) and C-C motif chemokine ligand 2 (CCL2) in tumors and increased expression of TNC on CAFs. In the 67NR model, the impact of vitamin D3 on blood flow or CAFs and lung NFs was not observed despite changes in plasma and/or tumor tissue concentrations of osteopontin (OPN), CCL2, transforming growth factor-β, vascular endothelial growth factor, and FGF23. In healthy mice, divergent effects of vitamin D3 supplementation/deficiency were observed, which lead to the creation of various body microenvironments depending on the mouse strain. Tumors developing in such microenvironments themselves modified the microenvironments by producing, for example, higher concentrations of OPN and stromal-cell-derived factor 1 (4T1), which influences the response to vitamin D3 supplementation/deficiency and calcitriol administration.

Insecticide Resistance Status of Malaria Vectors in a Malarious Area, Southeast of Iran.

Malaria continues to be the main vector-borne disease in Iran. The endemic foci of malaria are in Sistan and Baluchistan Province, the borderline of Iran and Pakistan. By the year 2020 the program of the country is malaria elimination. The main vector control is using insecticide as Indoor Residual Spraying. The aim of the study was to evaluate the susceptibility of main malaria vectors to different insecticides recommended by WHO. All the insecticides papers supported by WHO and evaluation of insecticide resistance of Anopheles stephensi, Anopheles culicifacies, Anopheles superpictus to different chemical groups of imagicides including DDT 4%, malathion 5%, propoxur 01.%, lambdacyhalothrin 0.05%, deltamethrin 0.025% and permethrin 0.75% were followed by the WHO guideline. Results of the susceptibility test against different insecticides revealed that An. stephensi and An. culicifacies are resistant to DDT and susceptible to other insecticides. An. superpictus is susceptible to all groups of pesticides. Knowledge on insecticide resistance in target species is a basic requirement to guide insecticide use in malaria control programmes in local and global scales.

Indeterminate pulmonary subsolid nodules in patients with no history of cancer: growing prediction, CT pattern, and pathological diagnosis.

PURPOSE We aimed to evaluate and compare the growth patterns among pathological types of inde- terminate subsolid nodules in patients without a history of cancer as observed on computed tomography (CT). METHODS This retrospective study included 77 consecutive patients with 80 indeterminate subsolid nod- ules on unenhanced thin-section CT. Subsolid nodules were classified into 2 growth pattern groups based on volume: growth (n = 35) and non-growth (n = 42). According to the pathologi- cal diagnosis, subsolid nodules were further subdivided into 3 groups: adenocarcinoma in situ (growth, n = 8 vs. non-growth, n = 22), minimally invasive adenocarcinoma (n = 14 vs. n = 15), and invasive adenocarcinoma (n=13 vs. n=5). Kaplan-Meier and Cox proportional hazards regres- sion analyses were performed to identify the risk factors for subsolid nodules growth. The CT findings of the 35 subsolid nodules in the growth group were compared among the 3 pathologi- cal groups. RESULTS In the growth group, the overall mean volume doubling time and mass doubling time (MDT) were 811.5 days and 616.5 days, respectively. Patient's age (odds ratio=1.041, P=.045) and CT subtype of non-solid nodule and part-solid nodule (odds ratio=3.430, P=.002) could predict subsolid nodule growth. The baseline volume, mass, and mean CT value were larger in the inva- sive adenocarcinoma group than in the adenocarcinoma in situ group (all P < .01). The shortest volume doubling time was observed in the invasive adenocarcinoma group, followed by the minimally invasive adenocarcinoma group and the adenocarcinoma in situ group. A shorter mass doubling time was observed in the minimally invasive adenocarcinoma group than in the adenocarcinoma in situ group (all P < .02). CONCLUSION As age increases, the risk of pulmonary subsolid nodule growth increases by 4% each year, and part-solid nodules have a 3 times higher risk of growth compared to non-solid nodules in patients with no history of cancer. Subsolid nodules with more aggressive pathological charac- teristics grow at a faster rate.

Effects of Pre-Anesthesia Anxiety on Propofol Induction Dose in Cats.

Simple SummaryIn humans, the anxiety felt by patients before anesthesia reportedly increases the anesthetic requirements for anesthesia induction. Relieving the patient’s anxiety before anesthesia is necessary to prevent the excessive administration of anesthetic. However, no studies have been conducted on the effects of anxiety on anesthesia induction in cats. Therefore, we aimed to investigate the impact of the anxiety healthy cats felt before anesthesia on the propofol dose required for inducing anesthesia. We observed that significant behavioral changes due to anxiety did not increase the propofol dose requirements in the cats compared to those who experienced no anxiety. Furthermore, there was no difference in the maintenance and recovery from anesthesia between the two aforementioned conditions. In conclusion, anxiety due to changing circumstances did not change the propofol dose required for anesthesia induction in cats, despite significant changes in their behavior.In humans, peri-anesthesia anxiety reportedly increases the anesthetic requirements for anesthesia induction. However, no studies have been conducted on cats regarding the effects of anxiety on anesthesia induction or anesthetic-mediated physiological changes. Therefore, we intended to investigate the effect of pre-anesthesia anxiety in healthy cats on the propofol dose required for anesthesia induction, and its impact on behavioral and physiological evaluations. The cats were placed in either a calm (CAL) or tense (ANX) environment. We performed physiological and behavioral evaluations before and after each environmental acclimatization period. Anesthesia was induced using propofol. We recorded the total dose of propofol administered for each clinical sign observed during anesthesia induction. The post-acclimatization behavioral evaluation score was significantly higher in the ANX group than the pre-acclimatization score. However, there was no significant difference in the propofol dose required for each clinical sign in the ANX or CAL groups. There were also no significant differences in the physiological evaluations between the ANX and CAL groups. Therefore, pre-anesthesia anxiety felt by cats did not affect propofol-mediated anesthesia induction.

Randomised controlled trial of the effects of increased energy intake on menstrual recovery in exercising women with menstrual disturbances: the 'REFUEL' study.

Does increased daily energy intake lead to menstrual recovery in exercising women with oligomenorrhoea (Oligo) or amenorrhoea (Amen)? A modest increase in daily energy intake (330 ± 65 kcal/day; 18 ± 4%) is sufficient to induce menstrual recovery in exercising women with Oligo/Amen. Optimal energy availability is critical for normal reproductive function, but the magnitude of increased energy intake necessary for menstrual recovery in exercising women, along with the associated metabolic changes, is not known. The REFUEL study (trial # NCT00392873) is the first randomised controlled trial to assess the effectiveness of 12 months of increased energy intake on menstrual function in 76 exercising women with menstrual disturbances. Participants were randomised (block method) to increase energy intake 20-40% above baseline energy needs (Oligo/Amen + Cal, n = 40) or maintain energy intake (Oligo/Amen Control, n = 36). The study was performed from 2006 to 2014. Participants were Amen and Oligo exercising women (age = 21.0 ± 0.3 years, BMI = 20.8 ± 0.2 kg/m2, body fat = 24.7 ± 0.6%) recruited from two universities. Detailed assessment of menstrual function was performed using logs and measures of daily urinary ovarian steroids. Body composition and metabolic outcomes were assessed every 3 months. Using an intent-to-treat analysis, the Oligo/Amen + Cal group was more likely to experience menses during the intervention than the Oligo/Amen Control group (P = 0.002; hazard ratio [CI] = 1.91 [1.27, 2.89]). In the intent-to-treat analysis, the Oligo/Amen + Cal group demonstrated a greater increase in energy intake, body weight, percent body fat and total triiodothyronine (TT3) compared to the Oligo/Amen Control group (P < 0.05). In a subgroup analysis where n = 22 participants were excluded (ambiguous baseline menstrual cycle, insufficient time in intervention for menstrual recovery classification), 64% of the Oligo/Amen + Cal group exhibited improved menstrual function compared with 19% in the Oligo/Amen Control group (χ2, P = 0.001). While we had a greater than expected dropout rate for the 12-month intervention, it was comparable to other shorter interventions of 3-6 months in duration. Menstrual recovery defined herein does not account for quality of recovery. Expanding upon findings in shorter, non-randomised studies, a modest increase in daily energy intake (330 ± 65 kcal/day; 18 ± 4%) is sufficient to induce menstrual recovery in exercising women with Oligo/Amen. Improved metabolism, as demonstrated by a modest increase in body weight (4.9%), percent body fat (13%) and TT3 (16%), was associated with menstrual recovery. This research was supported by the U.S. Department of Defense: U.S. Army Medical Research and Material Command (Grant PR054531). Additional research assistance provided by the Penn State Clinical Research Center was supported by the National Center for Advancing Translation Sciences, National Institutes of Health, through Grant UL1 TR002014. M.P.O. was supported in part by the Loretta Anne Rogers Chair in Eating Disorders at University of Toronto and University Health Network. All authors report no conflict of interest. NCT00392873. October 2006. September 2006.

Evaluation of Carotid Artery Elasticity in Early Stage of Kawasaki Disease by Two-Dimensional Speckle Tracking Imaging: A Pilot Study.

Children with Kawasaki disease (KD) often develop impaired arterial function. The aim of the study was to assess the feasibility and efficacy of two-dimensional speckle tracking technique (2DSTI) for the evaluation of carotid artery elasticity in children with early-stage KD. Children with KD (n=97), age and sex-matched children with fever (n=18), and healthy controls (n=24) were included. Children with KD were subsequently divided into a coronary artery lesion group (CAL group, 27 cases) and a noncoronary artery lesion group (nCAL group, 70 cases) based on the results of echocardiography. The carotid circumferential peak strain (CCS) and carotid intima-media thickness (CIMT) for the children in each group were measured, and the laboratory indicators for each group were collected. The CCS of children with KD was lower than that of children with fever and healthy controls (P=.001 and .008), whereas CIMT was not significantly different among the groups. Moreover, the CCS of children in the CAL group was lower than that of children in the nCAL group and healthy controls (P=.001 and .000, respectively), whereas the CIMT of children in the CAL group was higher than that of children in the nCAL group (P=.014). In children with KD, CCS was negatively correlated with C-reactive protein (CRP) and alanine aminotransferase (ALT) (r=-.419, P=.001; and r=-.305, P=.003). However, CCS was negatively correlated with CRP (r=-.508, P=.007) but not ALT (r=-.176, P=.379) in children in the CAL group. CCS determined based on 2DSTI can reflect changes in the carotid artery elasticity function in the early stage of KD.

Vitamin D Metabolite Profile in Cholecalciferol- or Calcitriol-Supplemented Healthy and Mammary Gland Tumor-Bearing Mice.

To analyze if the prometastatic activity of calcitriol (active vitamin D3 metabolite), which was previously observed in a 4T1 breast cancer model, is also found in other breast cancers, and to assess the impact of various schemes of vitamin D supply, we used 4T1 and E0771 mouse metastatic and 67NR nonmetastatic cells in this study. BALB/c and C57BL/6 healthy and tumor-bearing mice were exposed to a control (1000 IU), low- (100 IU), and high- (5000 IU) vitamin D3 diets. Additionally, from day 7 of tumor transplantation, the 1000 and 100 IU groups were gavaged with calcitriol (+cal). After 8 weeks of feeding, plasma levels of 25(OH)D3, 24,25(OH)2D3, and 3-epi-25(OH)D3 were significantly lower in calcitriol-treated and vitamin D-deficient groups than in the control, whereas the levels of all metabolites were increased in the 5000 IU group. The ratio of 25(OH)D3:24,25(OH)2D3 was increased in both calcitriol-treated groups, whereas the ratio of 25(OH)D3:3-epi-25(OH)D3 was increased only in the 100 IU group but decreased in the 5000 IU group. In contrast to E0771, 4T1 lung metastasis was accelerated in all vitamin D-supplemented mice, as well as in the deficient group with an increased inflammatory response. 67NR tumor growth was transiently inhibited in the 1000 IU+cal group, but single metastases were observed in the 5000 and 100 IU groups. Based on the results, we conclude that various schemes of vitamin D supply and vitamin D deficiency led to similar metabolite profiles irrespective of the mice strain and tumor burden. However, depending on the type of breast cancer, different effects on tumor growth and metastasis were noticed.

A Nutritional Intervention In Exercising Women With Oligo/Amenorrhea Improved Energy Availability

Reduced energy availability (EA) suppresses reproduction and induces oligo/amenorrhea (O/A) in exercising women. Treatment goals are to increase energy intake (EI) to improve EA and restore menstrual function. PURPOSE: Determine if a nutritional intervention that increased EI in exercising women with O/A improved EA and energetic status. METHODS A 12 month randomized controlled trial designed to increase energy intake 20-30% above baseline energy needs in exercising women with O/A randomized participants into O/A+CAL (n=13) and Control (n=10) groups. EI and exercise energy expenditure (EEE) were assessed throughout the intervention. EA was calculated as EI-EEE/fat free mass (FFM). Fasting blood samples were collected to assess TT3 and leptin. Independent t-tests/Mann-Whitney U tests compared differences between groups and correlations were run between changes in energy and hormones. Data are mean ± SE. RESULTS Subjects were 22±1 yr, BMI 20.0±0.4 kg/m2. Menstrual recovery was observed and previously reported. There were no pre intervention differences in EI (1837±95 kcal/d), EEE (390±40 kcal/d), FFM (41.5±0.8 kg) or EA (37.2±2.7 kcal/kg FFM/d) between groups. During the intervention there were greater absolute and percent increases in EI in O/A+CAL vs Control (448±170 vs -117±147 kcal/d, p=0.02; 23.6±8.6 vs -4.6±8.4%, p=0.03). There were no differences between groups in absolute or percent change for EEE (133±46 kcal/d, p=0.07; 38.8±11.3%, p=0.10) or FFM (0.6±0.3kg, p=0.60; 1.6±0.7%, p=0.51). Absolute increase in EA was greater in O/A+CAL vs Control (10.8±5.5 vs -6.0±3.9 kcal/kg FFM/d, p=0.03), but no difference in percent change (29.2±13.4 vs -6.7±12.5%; p=0.07). Absolute and percent increase in fat mass (2.3±0.3 vs 0.7±0.5, p<0.01; 20.6±3.1 vs 5.4±3.2%, p<0.01) and body mass (3.2±0.3 vs 0.8±0.8, p<0.01; 6.1±0.7 vs 1.5±1.3%, p<0.01) were greater in O/A+CAL vs Control. O/A+CAL had larger absolute and percent increases in TT3 (10.3±5.0 vs -6.7±5.3 ng/dL, p=0.03; 17.2±7.4 vs -8.3±5.5%, p=0.02) and percent increase in leptin (85.1±24.0 vs -13.3±18.0%) vs Control. Changes in EI and EA were not associated with changes in hormonal markers. CONCLUSION An intervention designed to improve EI can be considered an effective nutritional intervention for managing oligo/amenorrhea in exercising women.

Hunter-Gatherer Occupation of the Central Colorado Plateau during the Pleistocene-Holocene Transition

The central Colorado Plateau contains an exceptional density of cultural resources. Historically, however, archaeological investigations have overlooked the late Pleistocene and early Holocene record of this region. As such, there is currently a biased understanding of the earliest human occupations and adaptations. The regional Paleoindian record is reviewed here to assess the nature of initial human occupation of the area. Projectile point typologies, toolstone selection, and site distributions are used to characterize the land use patterns used by the region's earliest inhabitants. Results suggest that as early as circa 13,000 cal BP Clovis groups were familiar with the lithic landscape and habitually made use of local materials. Subsequent Paleoindian populations maintained relatively consistent levels of occupation throughout the Pleistocene-Holocene transition. Finally, the presence of classic Southwest, High Plains, and Great Basin–related lithic technologies suggests that the central Colorado Plateau was part of early interregional land use strategies.

The effect of chronic cerebral hypoperfusion on the pathology of Alzheimer's disease: A positron emission tomography study in rats

Cerebrovascular disease is a potential risk factor for Alzheimer's disease (AD). Although acute cerebral hypoperfusion causes neuronal necrosis and infarction, chronic cerebral hypoperfusion induces apoptosis in neurons, but its effects on the cognitive impairment are not clear. The purpose of this study was to evaluate the effects of chronic cerebral hypoperfusion on AD pathology and cerebral glucose metabolism. A model of chronic cerebral hypoperfusion was established by ligating the common carotid arteries bilaterally in adult male rats (CAL group). Sham-operated rats underwent the same procedures without artery ligation (control group). At 12 weeks after ligation, expression levels of amyloid-β (Aβ) and hyperphosphorylated tau (p-tau), as well as the regional cerebral glucose metabolism, were evaluated using Western blots and positron emission tomography with fluorine-18 fluorodeoxyglucose, respectively. The expression levels of Aβ in the frontal cortex and hippocampus and of p-tau in the temporal cortex were significantly higher in the CAL group than those in the control group. The cerebral glucose metabolism of the amygdala, entorhinal cortex, and hippocampus was significantly decreased in the CAL group compared to that in the control. These results suggest that chronic cerebral hypoperfusion can induce AD pathology and may play a significant role in AD development.

Calvin F. Quate (1923-2019).

Nanoscience pioneer who invented advanced microscopes

Resveratrol and 1,25-dihydroxyvitamin D co-administration protects the heart against D-galactose-induced aging in rats: evaluation of serum and cardiac levels of klotho.

The current study investigates the cooperative cardioprotective effect of calcitriol (active form of vitamin D) combined with resveratrol in a rat model of D-galactose (D.gal)-induced aging. Male Wistar rats received resveratrol (D.gal + Res), calcitriol (D.gal + Cal), or a combination of them (D.gal + Res + Cal). Intact animals served as control (Ctl). Blood pressure (BP) was recorded by cannulation of the left carotid artery. Fibrosis and cell size were assessed by Masson's trichrome and hematoxylin-eosin staining, respectively. Cardiac and serum level of antiaging protein, klotho, was measured by ELISA assay method. Gene expression was evaluated by real-time RT-PCR. Biochemical tests were performed according to the standardized method. In D.gal + Res + Cal group, BP, heart weight-to-body weight ratio, and cardiomyocytes size decreased significantly compared with D-gal group. The cardiac transcription levels of catalase and superoxide dismutase 1 and 2 were upregulated in D.gal + Res + Cal compared to the D.gal group (P < 0.001, P < 0.05, P < 0.05, respectively). Increased level of malondialdehyde was observed in D.gal group (P < 0.01 vs. Ctl) which was normalized partially in D.gal + Res + Cal group (P < 0.05). Catalase and superoxide dismutase activity also increased in D.gal + Res + Cal group (P < 0.01 vs. D.gal). Cardiac Klotho, as the antiaging protein, remained unchanged at mRNA and protein levels among the experimental groups. The serum level of Klotho did not change significantly in D.gal group; however, in D.gal + Res + Cal group, serum klotho concentration was increased (P < 0.05 vs. D.gal). It could be concluded that co-administration of resveratrol and vitamin D protects the heart against aging-induced damage by the modulation of hemodynamic parameters and antioxidant status of the heart. Furthermore, increased serum level of klotho could be a novel mechanism for antiaging effects of resveratrol and vitamin D.

Morphofunctional status of parotid salivary glands in three-month-old rats with experimentally induced fetal macrosomia

The paper aims at studying the features of postnatal morphogenesis of parotid salivary glands acinar tissue in three-month-old rats which were born with experimentally induced fetal macrosomia. The aim of the study is to examine experimentally the features of postnatal morphogenesis of salivary glands in three-month-old rats born with macrosomia. Materials and methods . Somatometric data of the animals were examined at birth and after the experiment, which induced macrosomia of rat fetus with four different ways (high-calorie diet for young pregnant female specimens, high-calorie diet for mature pregnant female specimens, high-calorie diet and hypokinesia condition for young pregnant female specimens, increased placental-fetal blood supply on the background of normal living conditions for young pregnant female specimens). To make the morphological analysis we used the tissues of parotid glands of three-month-old rats. We examined absolute and relative values of salivary gland mass. Beside the histological tests we used Gallocianin-chrome alum technique to detect DNA and RNA, and PAS-reaction to detect glycoproteins. Using computer images we measured the squares of serocytes, evaluated the content of DNA in the cell nucleus, the content of RNA in cytoplasm and glycoproteins in saliva in ducts and gaps of acini. Results. When rats reached the age of three months macrosomia-induced specimens with stimulated intrauterine growth (group Aks ) had a tendency to increase their average body weight while macrosomia-induced descendants of rats in the group 3 (high-calorie died and “soft” hypokinesia (group Y+Hk+cal ) had a tendency to decrease their body mass. And macrosomia-induced descendants of rats in the group 1 (high-calorie diet during pregnancy, Y+cal group) and decsendants in group 2 (high-calorie diet for mature rats during pregnancy, M+cal group) had somatometric data similar to the control-group. The relative mass of parotid glands of three-month-old rats which were born with macrosomia appeared to be increased in all the groups, but top values were observed in groups Y+cal and Y+Hk+cal . Rats with macrosomia from the group Aks by the age of three month had salivary glands with hypoplasia of serocytes which were morphofunctionally more active and had signs of proteins and glycoproteins hypoproduction. Rats with macrosomia in the Y+cal group by the age of three months had the amount of serocytes commensurate to the data of the control group but serocytes were more morphofunctionally active and had signs of glycoproteins hyperprodution. Rats in the M+cal group at the age of three months had serocytes hyperplasia, but their mophofunctional condition was similar to the control group. And macrosomia-induced rats in the Y+Hk+cal group had signs of the increase of morphofunctional activity of serocytes and hyperproduction of glycoproteins. Conclusion. It has been found that specimens which were born with macrosomia at the age of three months showed changes in morphofunctional state of parotid salivary glands parenchyma and, therefore, changes in the salivary composition in ducts and gaps of acini.

Immediate and late effects of chronic stress in the testes of prepubertal and adult rats.

The objective of this study was to investigate the effects of chronic stress on the testes of prepubertal and adult rats and to evaluate whether any alterations could be reversed when stress induction is ended. Seventy-six male rats were assigned to eight groups depending on the type of treatment (control or stressed), the age at which stress was initiated (prepubertal or adult), and the time of evaluation (immediate or late). Stress stimuli were applied for 6 weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimulus were included in SP-I and SA-I groups, respectively. The late prepubertal (SP-L) and adult (SA-L) groups of stressed rats were evaluated 6 weeks after the last stress stimulus. Age-matched rats were used as controls (CP-I, CA-I, CP-L, and CA-L groups). Application of stress stimuli to rats in the SP-I group resulted in body weight and seminiferous tubule diameter reduction. The rats in the SA-I group also showed several functional (testosterone level and sperm parameter) and morphological (testicular weight and seminiferous tubule diameter) reductions. The rats in the SP-L group showed increased body weight and intertubular compartment volumetric and absolute densities and reduced tubular compartment volumetric density. The rats in the SA-L group presented only reduced sperm viability. Stress stimuli promoted changes in the rats in all the study groups. The testes of the adult rats were the most affected by chronic stress. However, the stressed adult rats recovered well from the testicular alterations.

Marked increase in urinary excretion of apolipoproteins in children with nephrolithiasis associated with hypercalciuria

Using a proteomic approach, we aimed to identify and compare the urinary excretion of proteins involved in lipid transport and metabolism in children with kidney stones and hypercalciuria (CAL), hypocitraturia (CIT), and normal metabolic work-up (NM), and in healthy controls (HCs). Additionally, we aimed to confirm these results using ELISA, and to examine the relationship between the urinary excretion of selected proteins with demographic, dietary, blood, and urinary parameters. Prospective, controlled, pilot study of pooled urine from CAL, CIT, and NM versus age- and gender-matched HCs, using liquid chromatography-mass spectrometry. Relative protein abundance was estimated using spectral counting. Results were confirmed by ELISA performed on individual samples. Of the 1,813 proteins identified, 230 met the above criteria. Of those, 5 proteins (apolipoprotein A-II [APOA2]; apolipoprotein A-IV [APOA4]; apolipoprotein C-III [APOA3]; fatty acid-binding protein, liver [FABPL]; fatty acid-binding protein, adipocyte [FABP4]) involved in lipid metabolism and transport were found in the CAL group, with significant differences compared with HCs. ELISA analysis indicated statistically significant differences in the urinary excretion of APOC3, APOA4, and FABPL in the CAL group compared with HCs. Twenty-four-hour urinary calcium excretion correlated significantly with concentrations of ApoC3 (r = 0.77, p < 0.001), and FABPL (r = 0.80, p = 0.005). We provide proteomic data showing increased urinary excretion of lipid metabolism/transport-related proteins in children with kidney stones and hypercalciuria. These findings suggest that abnormalities in lipid metabolism might play a role in kidney stone formation.