table_1_miR-494 Contributes to Estrogen Protection of Cardiomyocytes Against Oxidative Stress via Targeting (NF-κB) Repressing Factor.xls

<p>Oxidative stress plays a pivotal role in the initiation and progression of cardiac diseases. Estrogens have been demonstrated to exert pleiotropic cardioprotective effects, among which antioxidative stress is one of the key effects linking estrogens to cardioprotection. By using a microRNAs (miRs) microarray screening approach, we discovered an increase in miR-494, which is known to exert cardioprotective effects, in estrogen-treated cardiomyocytes. We hypothesized that the upregulation of miR-494 might contribute to estrogen-mediated cardioprotection against oxidative stress. We found that E₂ stimulates miR-494 expression via ERα in both cardiomyocytes and the myocardium of female mice. The miR-494 inhibitor attenuated the protective effect of 17β-estradiol (E₂) against oxidative stress-induced injury in cardiomyocytes. By contrast, the miR-494 mimic protected cardiomyocytes against oxidative stress-induced cardiomyocyte injury. Using real-time PCR, western blot and dual-luciferase reporter gene analyses, we identified nuclear factor kappa B (NF-κB) repressing factor (NKRF) as the miR-494 target in cardiomyocytes. E₂ was found to inhibit NKRF, thus activating NF-κB through a miR-494-dependent mechanism. In addition, the protective effects of E₂ and miR-494 against oxidative stress in cardiomyocytes were eliminated by the NF-κB inhibitor. In summary, this study demonstrates for the first time that estrogen inhibits NKRF expression through ERα-mediated upregulation of miR-494 in cardiomyocytes, leading to the activation of NF-κB, which in turn results in an increase in antioxidative defense. ERα-mediated upregulation of miR-494 may contribute to estrogen protection of cardiomyocytes against oxidative stress.</p

CBS and CSE expression in pregnant myometrium before and after onset of labour.

<p>Myometrial tissues were obtained from pregnant women at term before the onset of labour (TNL, n = 10) or during active labour (TL, n = 10) A, the RT-PCR products of CBS and CSE. B, cumulative data of CBS and CSE mRNA levels in TNL and TL myometrium samples. The mRNA levels of CBS and CSE were determined by quantitative real-time RT-PCR as described in Material and Methods. Data were presented as mean±SEM. <i>*P</i><0.05, <i>**P</i><0.01 vs TNL. C, Western blot analysis of CBS and CSE in human pregnant myometrium. D, semiquantitation of Western blot signals of CBS and CSE in TNL and TL myometrium biopsies. Representative protein bands for CBS and CSE were on the top of histogram. Data were presented as mean±SEM.. <i>**P</i><0.01 vs TNL.</p

The effects of D-cysteine and taurine on myometrial contractility.

<p>Human myometrium strips were obtained from TNL patients (n = 4) undergoing elective cesarean section. Cumulative increases in D-cysteine (10⁻⁷–10⁻²mol/L) and taurine (10⁻⁷–10⁻²mol/L) were applied to myometrial strips. Spontaneous contractions and baseline muscle tension of myometrial strips were recorded. A&B, representative recording of contractions of myometrium strip treated with D-cysteine (A) or taurine (B). C,D&E, cumulative data of amplitude (C), frequency (D) and baseline tone (E). Data of amplitude and frequency were presented as mean (SEM) percentage of the results obtained before any drug application for each individual strip (control). For baseline muscle tone, the control was set as “0 ”. Data were presented as mean±SEM.</p

The expression of CBS and CSE in pregnant human myometrium.

<p>A&C shows representative sections for positive staining for CBS (A) and CSE (C). B&D shows the negative control sections stained with CBS preabsorption antibody (B) and CSE preabsorption antibody (D). E&F, negative control sections, the primary antibody was substituted by normal IgG(E) and PBS(F). Arrow: positive staining in myometrium smooth muscle cells. Original magnification ×400.</p

Effects of the K_ATP channel inhibitor on L-cysteine-induced changes in contractility of myometrium.

<p>K_ATP channel inhibitor glibenclamide (10⁻⁵mol/L) was applied to myometrial strips before addition of L-cysteine (10⁻³M). A&B, representative recording of nonlabouring(A) and labouring (B) myometrial strips were treated with L-cysteine. C&D, representative recording of nonlabouring(C) and labouring (D) myometrial strips treated with glibenclamide and L-cysteine. E,F&G, cumulative data of amplitude (E), frequency (F) and baseline tone (G). Data of amplitude and frequency were presented as mean (SEM) percentage of the results obtained before any drug application for each individual strip (control). For baseline muscle tone, the control was set as “0 ”. Data were presented as mean±SEM (TNL, n = 4; TL, n = 4). *<i>P</i><0.05;**<i>P</i><0.01 vs control. #<i>P</i><0.05; ##<i>P</i><0.01 vs L-cys. L-cys: L-cysteine.</p

H₂S production rate in human TNL and TL myometrium.

<p>A, Representative traces of H₂S production in the homogenate of myometrium tissues. H₂S production was initiated by the addition of L-cysteine and PLP. There were two control traces of buffered solutions without tissue or with heat-killed tissue containing L-cysteine and PLP, indicating no spontaneous H₂S production. B, Representative traces of AOAA and PAG blocking H2S production by the addition of L-cysteine in the homogenate of myometrium tissues. C, cumulative data of H2S production rate in TNL (n = 13) and TL myometrium (n = 6). Data were presented as mean?SEM. **P<0.01 vs TNL. L-Cys: L-cysteine.</p

Neuroanatomical site overlap.

Brain metastases are a frequent occurrence in neuropathology practices. The literature on their neuroanatomical location is frequently derived from radiological analyses. This work examines brain metastases through the lens of pathology specimens. All brain surgical pathology reports for cases accessioned 2011–2020 were retrieved from a laboratory. Specimens were classified by neuroanatomical location, diagnosis and diagnostic category with a hierarchical free text string-matching algorithm (HFTSMA) and also subsequently audited. All reports classified as probable metastasis were reviewed by a pathologist. The provided history was compared to the final categorization by a pathologist. The cohort had 4,625 cases. The HFTSMA identified 854 cases (including metastases from a definite primary, metastases from primary not known and improperly classified cases). 514/854 cases had one definite primary site per algorithm and on report review 538/854 cases were confirmed as such. The 538 cases originated from 511 patients. Primaries from breast, gynecologic tract, and gastrointestinal tract not otherwise specified were most frequently found in the cerebellum. Kidney metastases were most frequently found in the occipital lobe. Lung, metastatic melanoma and colorectal primaries were most commonly found in the frontal lobe. The provided clinical history predicted the primary in 206 cases (40.3%), was discordant in 17 cases (3.3%) and non-contributory in 280 cases (54.8%). The observed distribution of the metastatic tumours in the brain is dependent on the primary site. In the majority (54.8%) of cases, the provided clinical history was non-contributory; this suggests surgeon-pathologist communication may have the potential for optimization.</div

S2 File -

Brain metastases are a frequent occurrence in neuropathology practices. The literature on their neuroanatomical location is frequently derived from radiological analyses. This work examines brain metastases through the lens of pathology specimens. All brain surgical pathology reports for cases accessioned 2011–2020 were retrieved from a laboratory. Specimens were classified by neuroanatomical location, diagnosis and diagnostic category with a hierarchical free text string-matching algorithm (HFTSMA) and also subsequently audited. All reports classified as probable metastasis were reviewed by a pathologist. The provided history was compared to the final categorization by a pathologist. The cohort had 4,625 cases. The HFTSMA identified 854 cases (including metastases from a definite primary, metastases from primary not known and improperly classified cases). 514/854 cases had one definite primary site per algorithm and on report review 538/854 cases were confirmed as such. The 538 cases originated from 511 patients. Primaries from breast, gynecologic tract, and gastrointestinal tract not otherwise specified were most frequently found in the cerebellum. Kidney metastases were most frequently found in the occipital lobe. Lung, metastatic melanoma and colorectal primaries were most commonly found in the frontal lobe. The provided clinical history predicted the primary in 206 cases (40.3%), was discordant in 17 cases (3.3%) and non-contributory in 280 cases (54.8%). The observed distribution of the metastatic tumours in the brain is dependent on the primary site. In the majority (54.8%) of cases, the provided clinical history was non-contributory; this suggests surgeon-pathologist communication may have the potential for optimization.</div

Distribution of metastatic tumours by primary site (Pathobiologic Perspective).

Distribution of metastatic tumours by primary site (Pathobiologic Perspective).</p

Patients by primary site and Lesions by primary site and location.

Patients by primary site and Lesions by primary site and location.</p