Differential effects of chronic stress in young-adult and old female mice: cognitive-behavioral manifestations and neurobiological correlates.

Stress-related psychopathology is highly prevalent among elderly individuals and is associated with detrimental effects on mood, appetite and cognition. Conversely, under certain circumstances repeated mild-to-moderate stressors have been shown to enhance cognitive performance in rodents and exert stress-inoculating effects in humans. As most stress-related favorable outcomes have been reported in adolescence and young-adulthood, this apparent disparity could result from fundamental differences in how aging organisms respond to stress. Furthermore, given prominent age-related alterations in sex hormones, the effect of chronic stress in aging females remains a highly relevant yet little studied issue. In the present study, female C57BL/6 mice aged 3 (young-adult) and 20-23 (old) months were subjected to 8 weeks of chronic unpredictable stress (CUS). Behavioral outcomes were measured during the last 3 weeks of the CUS protocol, followed by brain dissection for histological and molecular end points. We found that in young-adult female mice, CUS resulted in decreased anxiety-like behavior and enhanced cognitive performance, whereas in old female mice it led to weight loss, dysregulated locomotion and memory impairment. These phenotypes were paralleled by differential changes in the expression of hypothalamic insulin and melanocortin-4 receptors and were consistent with an age-dependent reduction in the dynamic range of stress-related changes in the hippocampal transcriptome. Supported by an integrated microRNA (miRNA)-mRNA expression analysis, the present study proposes that, when confronted with ongoing stress, neuroprotective mechanisms involving the upregulation of neurogenesis, Wnt signaling and miR-375 can be harnessed more effectively during young-adulthood. Conversely, we suggest that aging alters the pattern of immune activation elicited by stress. Ultimately, interventions that modulate these processes could reduce the burden of stress-related psychopathology in late life.

Implementing diffusion-weighted MRI for body imaging in prospective multicentre trials: current considerations and future perspectives.

For body imaging, diffusion-weighted MRI may be used for tumour detection, staging, prognostic information, assessing response and follow-up. Disease detection and staging involve qualitative, subjective assessment of images, whereas for prognosis, progression or response, quantitative evaluation of the apparent diffusion coefficient (ADC) is required. Validation and qualification of ADC in multicentre trials involves examination of i) technical performance to determine biomarker bias and reproducibility and ii) biological performance to interrogate a specific aspect of biology or to forecast outcome. Unfortunately, the variety of acquisition and analysis methodologies employed at different centres make ADC values non-comparable between them. This invalidates implementation in multicentre trials and limits utility of ADC as a biomarker. This article reviews the factors contributing to ADC variability in terms of data acquisition and analysis. Hardware and software considerations are discussed when implementing standardised protocols across multi-vendor platforms together with methods for quality assurance and quality control. Processes of data collection, archiving, curation, analysis, central reading and handling incidental findings are considered in the conduct of multicentre trials. Data protection and good clinical practice are essential prerequisites. Developing international consensus of procedures is critical to successful validation if ADC is to become a useful biomarker in oncology. KEY POINTS: • Standardised acquisition/analysis allows quantification of imaging biomarkers in multicentre trials. • Establishing "precision" of the measurement in the multicentre context is essential. • A repository with traceable data of known provenance promotes further research.

The fetal safety of clomiphene citrate: a population-based retrospective cohort study.

OBJECTIVE: To evaluate whether exposure to clomiphene citrate (CC) for ovulation induction is associated with major malformations overall or with specific fetal anomalies. DESIGN: We conducted a population-based retrospective cohort study. Exposure was defined as CC dispension from 2 months before conception through the first month of pregnancy. SETTINGS: Four databases were combined: medication, birth, hospitalization, and terminations of pregnancy. POPULATION: The study included all women in southern Israel who gave birth or underwent termination of pregnancy at Soroka Medical Center, from 1998 to 2009. METHODS: The rates of major malformations overall and six different subcategories of anomalies were evaluated. The crude odds ratio (OR) was calculated with a 95% confidence interval (95% CI). Subsequently the adjusted odds ratio (aOR) was calculated using multiple logistic regression models controlling for maternal age, pre-pregnancy diabetes, parity, ethnicity, the calendar year in which the birth/termination of pregnancy took place, smoking, and the use of gonadotropins and progesterone. MAIN OUTCOME MEASURES: Major malformations overall and specific fetal malformations by organ systems. RESULTS: Of 114 961 pregnant women, 1872 were exposed to CC. No association was detected between exposure to CC and rates of major malformations overall (aOR 1.08, 95% CI 0.88-1.32) or rates of subcategories of malformations. Exposure was not associated with anencephaly (aOR 2.27, 95% CI 0.44-11.71) or oesophageal atresia (aOR 3.681, 95% CI, 0.65-20.76). CONCLUSIONS: In this large population-based retrospective cohort study, exposure to CC was not associated with an increased risk of either rates of major malformations overall or rates of specific malformations. TWEETABLE ABSTRACT: An observational study: no increased risk for fetal malformations following exposure to clomiphene citrate.

Squeezing and entanglement in a Bose-Einstein condensate.

Entanglement, a key feature of quantum mechanics, is a resource that allows the improvement of precision measurements beyond the conventional bound attainable by classical means. This results in the standard quantum limit, which is reached in today's best available sensors of various quantities such as time and position. Many of these sensors are interferometers in which the standard quantum limit can be overcome by using quantum-entangled states (in particular spin squeezed states) at the two input ports. Bose-Einstein condensates of ultracold atoms are considered good candidates to provide such states involving a large number of particles. Here we demonstrate spin squeezed states suitable for atomic interferometry by splitting a condensate into a few parts using a lattice potential. Site-resolved detection of the atoms allows the measurement of the atom number difference and relative phase, which are conjugate variables. The observed fluctuations imply entanglement between the particles, a resource that would allow a precision gain of 3.8 dB over the standard quantum limit for interferometric measurements.

SPECT-CT in total hip arthroplasty.

Single photon emission computed tomography combined with computed tomography (SPECT-CT), which combines functional and anatomical imaging, provides superior spatial localization to plain radiography and is more tolerant to metallic artefacts than conventional imaging such as magnetic resonance imaging (MRI). It is increasingly used in musculoskeletal imaging to enable accurate anatomical localization of increased tracer uptake, and is particularly useful in assessing metal prosthesis and the surrounding bone following total hip arthroplasty (THA). In addition to detecting complications of THA, SPECT-CT enables multiplanar reconstruction and manipulation of imaging data, which may aid surgical planning. SPECT-CT is an important adjunct to conventional imaging in the management of post-THA complications. It is vital that radiologists are able to identify the specific features of different complications and use this novel imaging technique to guide management. In this article, the use of SPECT-CT to follow post-THA complications, including aseptic loosening, periprosthetic infection, histiocytic reactions, periprosthetic fractures, polyethylene wear, and pseudotumour formation, will be reviewed.

Does the extent of neck surgery based on preoperative calcitonin level influence survival in medullary thyroid carcinoma: a retrospective tertiary centre experience.

INTRODUCTION: Medullary thyroid carcinoma (MTC) is a rare thyroid cancer arising from the calcitonin-secreting parafollicular cells. Previous studies suggested a preoperative calcitonin level >200ng/l is an indication for prophylactic lateral neck dissection (LND) to remove micrometastases and improve survival outcomes. METHODS: This retrospective single-centre study assessed the efficacy of preoperative calcitonin levels as a marker for determining need for prophylactic LND in MTC. Data were obtained on demographics, preoperative calcitonin levels, size and laterality of index tumour, type of neck dissection (central neck dissection (CND), LND), nodes removed, levels with involved nodes, number of nodes histologically involved, mortality, adjuvant therapy and locoregional recurrence. RESULTS: A total of 33 patients were identified from St George's University Hospitals NHS Foundation Trust between 1 January 2001 and 19 August 2021; 8 were excluded for data quality issues. Of the 18 classified with a high preoperative calcitonin level (>200ng/l), 10 (56%) had a LND and 8 (44%) had a CND. In the low-calcitonin group, three (43%) patients had a CND only and four (57%) had a LND. There was no difference in absolute or disease-free survival between the low and high groups (p=0.960, p=0.817), or between those who had a CND and LND in the high-calcitonin group (p=0.607, hazard ratio (HR) 0.55; p=0.129, HR 8.78). CONCLUSION: There was no statistically significant difference in outcomes between high and low calcitonin groups. A selective approach to performing LND in MTC patients based on clinical and imaging findings suggesting disease presence in the lateral neck should be explored further.

Unraveling the molecular basis of cannabidiolic acid methyl Ester's anti-depressive effects in a rat model of treatment-resistant depression.

Major depressive disorder (MDD) stands as a significant cause of disability globally. Cannabidiolic Acid-Methyl Ester (CBDA-ME) (EPM-301, HU-580), a derivative of Cannabidiol, demonstrates immediate antidepressant-like effects, yet it has undergone only minimal evaluation in psychopharmacology. Our goal was to investigate the behavioral and potential molecular mechanisms associated with the chronic oral administration of this compound in the Wistar Kyoto (WKY) genetic model of treatment-resistant depression. Male WKY rats were subjected to behavioral assessments before and after receiving chronic (14-day) oral doses of CBDA-ME (0.5 mg/kg), 15 mg/kg of imipramine or vehicle. At the end of the study, plasma corticosterone levels and mRNA expression of various genes in the medial Prefrontal Cortex and Hippocampus were measured. Behavioral outcomes from CBDA-ME treatment indicated an antidepressant-like effect similar to imipramine, as oral ingestion reduced immobility and increased swimming duration in the Forced Swim Test. Neither treatment influenced locomotion in the Open Field Test nor preference in the Saccharin Preference Test. The behavioral impact in WKY rats coincided with reduced corticosterone serum levels, upregulated mRNA expression of Cannabinoid receptor 1, Fatty Acid Amide Hydrolase, and Corticotropin-Releasing Hormone Receptor 1, alongside downregulation of the Serotonin Transporter in the hippocampus. Additionally, there was an upregulation of CB1 mRNA expression and downregulation of Brain-Derived Neurotrophic Factor in the mPFC. These findings contribute to our limited understanding of the antidepressant effects of CBDA-ME and shed light on its potential psychopharmacological mechanisms. This discovery opens up possibilities for utilizing cannabinoids in the treatment of major depressive disorder and related conditions.

Optimising the acquisition and retention of heat acclimation.

Heat acclimation (HA) often starts in a moderately hot environment to prevent thermal overload and stops immediately prior to athletic activities. The aims of this study were (1) to establish whether acclimation to a moderately hot climate is sufficient to provide full acclimation for extreme heat and (2) to investigate the physiological responses to heat stress during the HA decay period. 15 male subjects exercised for 9 consecutive days at 26° C Wet Bulb Globe Temperature (WBGT) and 3 days at 32° C WBGT on a cycle ergometer for up to 2 h per day and repeated the exercise 3, 7 and 18 days later in 26° C WBGT. Rectal temperature (T (re)) and heart rate (HR) were measured during 60 min of steady state exercise (∼45% of maximum oxygen uptake). During days 1-9, end-exercise T (re) was reduced from 38.7±0.1 to a plateau of 38.2±0.1° C (p<0.05), HR was reduced from 156±10 to 131±11 bpm (p<0.05). No changes in HR and T (re) occurred during the 3 days in the very hot environment. However, T (re) during rest and exercise were significantly lower by 0.4-0.5° C after HA compared with day 9, suggesting that heat acclimation did not decay but resulted in further favourable adaptations.

Terminal complement proteins C5b-9 release basic fibroblast growth factor and platelet-derived growth factor from endothelial cells.

Interactions between endothelium and vascular smooth muscle cells play a major role in the biology of the blood vessel wall. Growth factors released from endothelial cells control in part the normal and pathological proliferation of vascular smooth muscle cells. Endothelial deposits of C5b-9 proteins, the membrane attack complex of complement (MAC), have been found in a variety of pathological tissues in which cell proliferation is an early characteristic abnormality, including atherosclerosis. We have explored a possible bridging role for terminal complement C5b-9 proteins in eliciting focal signals for cell proliferation by releasing growth factors from endothelial cells. We found that both bovine aortic and human umbilical vein cells respond to the MAC by releasing basic fibroblast growth factor and platelet-derived growth factor. These mitogens stimulate DNA synthesis in Swiss 3T3, vascular smooth muscle, and glomerular mesangial cells. Based on these findings, we propose that complement-induced release of mitogens from endothelial cells is a novel pathogenic mechanism for proliferative disorders.

The role of specific antibody in alternative complement pathway-mediated opsonophagocytosis of type III, group B Streptococcus.

The native capsular polysaccharide antigen of type III, group B Streptococcus contains a terminal sialic acid residue on each repeating unit that masks all end-group galactopyranose residues and prevents alternative pathway complement activation by adult human sera in the absence of type-specific antibody. The critical role of the sialic acid residues in allowing the organism to evade activating the alternative complement pathway was shown when neuraminidase treatment of the organism converted the bacteria to activators of the alternative pathway as assessed in agammaglobulinemic serum. The requirement for specific antibody in permitting alternative pathway activation by the fully sialated bacteria was shown when sera that contained low levels of specific antibody failed to activate this pathway, and when prior absorption of serum that contained higher type-specific antibody levels with the capsular antigen failed to activate this pathway. The use of C2-deficient sera showed that the calssical pathway was not required for antibody-dependent alternative pathway activation. The use of isotonic, pH 7.5, veronal-NaCl buffer that contained 1% gelatin and that was supplemented to 4 mM Mg++ and 16 mM EGTA and adjusted to pH 7.5 (MgEGTA) ruled out the participation of the C1-bypass pathway. The presence of sialic acid on the bacterial surface is one means of evading an important mechanism of natural immunity, namely activation of complement by the alternative pathway. Only specific antibody, i.e., acquired immunity, can overcome this virulence factor.

Human eosinophils express CD4 protein and bind human immunodeficiency virus 1 gp120.

The CD4 glycoprotein, expressed on leukocytes belonging to subsets of T lymphocytes and to cells of monocyte/macrophage lineage, participates in the functioning of T cells and serves as a receptor for HIV-1 and HIV-2. Human eosinophils, a class of granulocytic leukocytes, have been found to express CD4. With anti-CD4 mAbs CD4 was demonstrable on eosinophils from both normal and eosinophilic donors. Eosinophils synthesized a 55-kD CD4 polypeptide immunoprecipitable with two anti-CD4 mAbs. Eosinophil CD4 bound HIV-1 gp120 as assessed by competition for anti-OKT4A, but not anti-OKT4, mAb binding. Eosinophils, normally rich in gastrointestinal and genitourinary tract tissues, increase in numbers in patients with metazoan parasitic infections. In these sites and diseases, CD4 expression by eosinophils may be pertinent to their immunologic functions and could make these cells susceptible to HIV infection.

Complement receptor 1/CD35 is a receptor for mannan-binding lectin.

Mannan-binding lectin (MBL), a member of the collectin family, is known to have opsonic function, although identification of its cellular receptor has been elusive. Complement C1q, which is homologous to MBL, binds to complement receptor 1 (CR1/CD35), and thus we investigated whether CR1 also functions as the MBL receptor. Radioiodinated MBL bound to recombinant soluble CR1 (sCR1) that had been immobilized on plastic with an apparent equilibrium dissociation constant of 5 nM. N-acetyl-d-glucosamine did not inhibit sCR1-MBL binding, indicating that the carbohydrate binding site of MBL is not involved in binding CR1. C1q inhibited MBL binding to immobilized sCR1, suggesting that MBL and C1q might bind to the same or adjacent sites on CR1. MBL binding to polymorphonuclear leukocytes (PMNs) was associated positively with changes in CR1 expression induced by phorbol myristate acetate. Finally, CR1 mediated the adhesion of human erythrocytes to immobilized MBL and functioned as a phagocytic receptor on PMNs for MBL-immunoglobulin G opsonized bacteria. Thus, MBL binds to both recombinant sCR1 and cellular CR1, which supports the role of CR1 as a cellular receptor for the collectin MBL.

Rare case of venous tumour thrombus from acinic cell carcinoma of the parotid gland.

Head and neck tumour thrombus is a rare pathology and at present there are no reported cases of tumour thrombus secondary to acinic cell carcinoma of the parotid gland. We report a case of an 81-year-old man with an acinic cell carcinoma of the left parotid and an intravenous tumour thrombus extending from the retromandibular vein into the internal jugular vein. This case also highlights the importance of radiological imaging in the management of tumour thrombus.

Si-C(sp3) bond activation through oxidative addition at a Rh(i) centre.

An easy, direct and room temperature silicon-carbon bond activation is reported. The reaction of [RhCl(coe)2]2 with the silane Si(Me)2(o-C6H4SMe)2 in the presence of an halide extractor provokes a Si-CH3 bond cleavage yielding a cationic silyl-methyl-Rh(iii). In contrast, if the reaction is performed using the Rh(i) bis-alkene dimers, [RhCl(cod)]2 or [RhCl(nbd)]2, the Si-CH3 bond activation does not occur.

Amino acid sequence of mouse tenascin and differential expression of two tenascin isoforms during embryogenesis.

We have isolated cDNA clones for mouse tenascin and analyzed expression of tenascin mRNAs during embryonic development of the kidney and gut. The deduced amino acid sequence of the mouse tenascin cDNAs shows a modular structure of repeats similar to chicken and human tenascin. In mouse there are 14.5 cysteine-rich repeats with similarity to the EGF repeat, followed by several repeats with similarity to the type III repeat of fibronectin. A longer variant contains 13 fibronectin type III repeats, whereas a shorter splice variant of mouse tenascin lacks the 5 type III repeats that occur directly after the fifth repeat in the longer variant. Contrary to the chicken and human sequences, mouse tenascin does not contain an RGD sequence in the third type III repeat implicated in cell attachment, or in any other positions. In Northern hybridizations to RNA from primary embryonic fibroblasts, the cDNA clone M 20/1 detects two mRNAs with sizes close to 6 and 8 kb. This, and the other data presented here suggest that the two major mouse tenascin polypeptides arise through an alternative RNA splicing. The two major mRNAs are differentially expressed during development. The 8-kb mRNA is more prominent than the 6-kb mRNA throughout prenatal kidney development, but during postnatal development the ratio of the two mRNAs changes. A different expression pattern is seen in the developing gut where the 6-kb mRNA predominates during embryogenesis with the 8-kb mRNA appearing later. The mRNA data of the developing gut correspond with previous protein data, which showed that the shorter Mr 210,000 polypeptide predominates during earlier developmental stages and the larger Mr 260,000 polypeptide appears later in the embryonic gut (Aufderheide, E., and P. Ekblom. 1988. J. Cell Biol. 107:2341-2349).

Endogenous cholecystokinin reduces feeding in young rats.

The hypothesis that endogenous cholecystokinin (CCK) released from the small intestine during feeding causes satiety was tested in rat pups, 9 to 12 days old. Intragastric administration of soybean trypsin inhibitor, a procedure that releases CCK from the small intestine, decreased the subsequent intake of a test meal. This effect was reversed by prior treatment with MK-329, a selective antagonist of CCK at alimentary-type CCK (CCK-A) receptors. Thus, endogenous, small intestinal CCK can cause satiety in the neonatal rat and this effect involves CCK-A receptors.

Divergent maternal behavioral patterns in two genetic animal models of depression.

Maternal behavior was examined in Flinders Sensitive-Line (FSL) and Wistar-Kyoto (WKY) rats, two different genetic animal models of depression. Behavioral patterns were assessed by undisturbed observations in the nest [Post-Partum Days (PPD) 4 and 9] and post-disturbance observations ("retrieval tests") on PPD 10. Litters were randomly allocated to a mild chronic-stress condition (limiting available bedding between PPD 2 and 9) or a standard rearing condition. The findings indicated that FSL dams did not differ from control dams in the undisturbed observations. However, in the post-disturbance observations FSL dams exhibited less pup-directed behaviors, a shorter latency to first pup carrying/retrieval and more self-directed behaviors than controls (the latter effect only in dams' interaction with whole litter). In contrast, WKY dams performed more pup-directed activities (e.g., nursing and licking) and less self-directed activities in both the undisturbed and post-disturbance observations (in both dams' interaction with single-pup and with the whole-litter) compared to controls. Accordingly, WKY dams exhibited a shorter latency for first pup-licking bout (in both post-disturbance observations). The early life mild chronic-stress used in the study ('limited-bedding') had a minor effect on the dams' behavior. Overall, the study investigated for the first time the maternal behavior of WKY dams and suggests that these dams show an almost opposite behavioral pattern to that of FSL dams. The results are discussed with regard to earlier findings in the FSL strain and behavioral patterns documented in depressed human mothers.

Adaptation to lactation in OLETF rats lacking CCK-1 receptors: body weight, fat tissues, leptin and oxytocin.

OBJECTIVE: To understand the adaptation to lactation of obese rats, by studying the interplay among the gut hormone cholecystokinin (CCK), the adiposity hormone leptin and the affiliation hormone oxytocin in modulating body mass and fat storage. DESIGN: Strain differences were examined between Otsuka Long Evans Tokushima Fatty (OLETF) rats lacking expression of functional CCK-1 receptors and Long Evans Tokushima Otsuka (LETO) controls, tested as nulliparous dams, at the 7 and 15th lactation day, at weaning (lactation day 22) or 8 weeks postweaning. MEASUREMENTS: We measured body mass, fat pads (brown, retroperitoneal and inguinal) and inguinal adipocytes. Plasma levels of leptin and oxytocin were determined. RESULTS: Fat depots of LETO female rats were larger during lactation compared to the levels found in postweaning and nulliparous female rats. LETO female rats gained weight and accumulated fat during pregnancy and lactation, returning to their normal fat levels postweaning. In contrast, OLETF female rats presented lower body weight and fat depots during the lactation period than nulliparous dams, and regained the weight and fat postweaning. Plasma leptin and oxytocin were highly correlated and followed the same pattern. OLETF leptin levels were highly correlated with fat depot and inguinal cell surface. No significant correlation was found for LETO parameters. CONCLUSIONS: Pregnancy and lactation are energy-consuming events, which naturally induce female rats to increase food intake and accumulate fat. When challenged by the demands of rapidly growing preobese OLETF pups, OLETF dams' fat stores are reduced to lean, LETO levels. During lactation, sensitivity of the oxytocinergic neurons descending from the paraventricular nuclei to the nucleus of the solitary tract to CCK is reduced. We theorized that this pathway is not available to OLETF female rats that lack functional CCK-1 receptors to mediate the signal. The current study contributes to the understanding of the female body's adaptation to lactation.

Withdrawal emotional-regulation in infant rats from genetic animal models of depression.

Children of depressed parents exhibit high rates of emotion-dysregulation, characterized by excessive withdrawal or approach strategies toward the mother in infancy. The understanding of factors affecting the establishment of these behavioral deficits is limited. The current study utilized two genetic animal models of depression, the Wistar-Kyoto (WKY) and Flinders Sensitive Line (FSL) rat strains. In addition, in order to assess the interactive effects of depressive vulnerability and exposure to early life stress, the subjects were raised either in a standard rearing condition or exposed to mild chronic-stress on postnatal days (PND) 2-9. On PND 10-11, an isolation test examined the pups' emotion-regulation. WKY pups produced less separation-induced ultrasonic vocalizations (USV) and proximity-seeking behaviors, compared to controls. In addition, WKY pups did not show the expected potentiation effect that was evident in control pups (an increase in USV and pivoting behavior after a short reunion with the dam). FSL pups exhibited less proximity-seeking behaviors compared to controls while showing levels of USV, potentiation of USV, and change in proximity-seeking behaviors that were similar to controls. No differences between the strains were found in self-grooming. The early life chronic-stress paradigm had no effect on the behaviors of the pups, indicating either stress-resilience or a limited effect of the paradigm. Overall, the results tentatively imply a tendency of the WKY and FSL pups towards withdrawal behavior instead of approach-behavior when regulating emotion, with a more pronounced pattern in WKY pups. This behavioral profile is reminiscent of avoidant attachment, a characteristic of many children of depressed parents.