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The Student Athlete in the Pandemic | A Tale of Two Americas
Posted On March 8, 2021 March 8, 2021 By Callum Buchan - Sports Editor
The United States has been the preferred location for many of Ireland's student athlete, looking to achieve their sporting ambitions whilst obtaining a degree, for the best of a century.
It was the path chosen by Olympic silver-medallist Sonia O'Sullivan and major-winning golfer Graeme McDowell, as well as thousands of other Irish people who took the opportunity to study in America's luxurious universities, often avoiding the exorbitant tuition fees, as a result of their athletic talent.
The current crop of scholarship students find themselves in a unique set of circumstances, in comparison with the vast swathes who preceded them, as they are competing and studying in the midst of a global pandemic. The College Tribune spoke to four Irish students on sports scholarships in the US to hear their perspective on life stateside as a student-athlete in the context of COVID-19.
Julie McCarthy. Photograph: Kristen Bradshaw
It was during the practice round of a tournament in Georgia when Julie McCarthy, a senior year Accounting major on a golf scholarship at Auburn University in Alabama, began hearing of COVID-19 hitting the US in March 2020. The tournament was subsequently cancelled and McCarthy's team travelled back to Auburn. "Then everything happened so fast, within a few days we heard that the university was shutting down and all lectures were going online," McCarthy said.
McCarthy flew back to her family in Swords in late March once any student-visas issues were sorted. Since returning to Auburn in August, in-person classes have been optional at half-capacity while the golf team has been in a bubble in which they are tested twice a week. Last semester, four of the team tested positive which disrupted plans for their first tournament. The team are advised to only socialise within the bubble and to avoid restaurants and other busy places at peak times.
McCarthy admitted that they "can still do things (socially), but it's on a different level now" compared to pre-COVID when, confirming the stereotype, much of the students' social events consisted of red cup parties and beer-pong tournaments. "I wish I could say differently but there was a lot of that!" Outside of these measures, McCarthy said things have been "pretty much normal here, absolutely everything is open". The 22-year-old will graduate next May as Auburn has given students an extra year back due to COVID.
The experience of Stephanie Ryan during the pandemic starkly contrasts that of McCarthy's. The 22-year-old student-athlete squash player at Drexel University in Philadelphia was in a packed bar for Erin Express in March. "It is basically St Patrick's Day celebrations," she said, "but for three weekends straight," with COVID barely being mentioned.
Soon after this, classes were all put online the week before Spring Break and Ryan was due to start a co-op in April with a financial services company on Wall Street. "In the space of two weeks it went from not hearing about restrictions to not knowing whether I could even get into New York," she said.
With this uncertainty, she made the early trip to the 'Big Apple' to stay with her brother until her own apartment opened up. Ryan worked remotely for six months which she found very tough at times as she was living alone in a new city during a lockdown, but said the company was very supportive. The once uber-busy NYC was empty which made Ryan feel anxious about going for long walks.
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In May, there were more reasons to feel unsafe as some protests turned to rioting and looting in Manhattan. "I thought I'll stay inside just to be careful," Ryan said. After Thanksgiving, Philadelphia became a high-level contagious city and has been in and out of lockdown since. "Wherever I was always seemed to be the worst hit county!". All of Ryan's Computer Science classes are taught online at Drexel and she is yet to play squash since last March despite testing protocols being put in place throughout the college. "The worst part of it is we've all worked really hard over the years, and now I probably won't play with my friends for my last year or even have a graduation," she said.
The Freshers' Perspective for a Student Athlete
"Coming over here, honestly, I didn't think I'd last". This is how Jack Casey, a 19-year-old originally from Newbridge, felt before making the move across the Atlantic to start a new chapter of his life at Keiser University, Miami on a football scholarship. However, there are many international students who are in the same situation as himself and he met a good group of friends who made the transition a lot easier than anticipated.
The left back has been a mainstay for the Keiser men's soccer team this season, who went unbeaten until the final of The Sun Conference. In October, the team experienced their only hiccup in relation to the pandemic as eight of the team tested positive for COVID, including Casey who experienced symptoms for the first three days, meaning all training and matches were cancelled for two weeks.
Jack Casey. Photograph: Milton Graham
The players must wear masks at all times indoors and are "tested every so often, but not all of the time", but other than that it is business as usual, according to Casey. His Sports Management lectures are in a mixed format, with in-person classes twice a week. The restrictions in Florida are "much more laid-back than Ireland," with even nightclubs being open in the 'Sunshine State'. Casey's friend group enjoy going to restaurants and the driving range in their free time and he said "It (COVID) hasn't negatively affected my social life too much, but I can see how it's affected people back home".
Hollywood has made us associate life in US colleges with frat parties, beer-pong and American football games in front of thousands. We certainly don't picture it starting with two weeks of quarantine which coincides with receiving your Leaving Cert results. However, this was Ali O'Dea's first experience at Elon University, North Carolina. "It was hard being alone and seeing all of your friends back home celebrating," O'Dea said.
Since this period, O'Dea has thoroughly enjoyed her tennis scholarship as everyone on her team is "super welcoming and like-minded". The college has done a lot to ensure the safety of athletes during the pandemic, as O'Dea is in a bubble with her teammates and must produce a negative antigen test three times a week in order to play matches.
She was deemed a close contact last semester which resulted in another period of isolation and any positive cases must go through cardiovascular testing before competing again. Similar to Auburn and Miami, O'Dea said that most places are open and there is yet to be a lockdown in Elon; however, there is a 10pm curfew in place. Within the bubble they "can do small bits of socialising but nothing like college expectations," O'Dea said. The majority of classes at Elon are in-person, but with the tennis team's packed schedule O'Dea often finds herself watching lectures through Zoom on the bus to and from matches.
Ali O'Dea. Photograph: Elon University
The effects of the pandemic on students in Ireland are apparent to all, there are few parts of the college life that have not been radically altered in the last year. Many students, who find themselves in the never ending Groundhog Day of lockdown where socialising is reduced to a three minute breakout room in a Zoom lecture, will look to some of these stories from America with envy.
However, the student experience in the States is far from a harmonious one as there is little consensus between states on restrictions. Political, cultural and economic polarisation in the United States have been rife over the last decade and the COVID-19 response has been no different. In many ways the differing circumstances of the student-athletes during the pandemic acts as a bellwether for this division.
Callum Buchan – Sports Writer
Tagged: America Soccer student athletes students students abroad Tennis
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Resveratrol alleviates lipopolysaccharide-induced inflammation in PC-12 cells and in rat model
Guiqi Zhang1,
Yi Liu1,
Lichen Xu1,
Chunhe Sha1,
Haibin Zhang1 &
Weibing Xu1
Spinal cord injury (SCI) remains a huge medical problem nowadays as there is no hospital providing the versatile strategies for repairing central nervous system and restoring function. Herein, we focused on PC-12 cells as an important research tool and studied the potential role of resveratrol (RSV) in inflammation induced by LPS.
RSV improved inflammatory injury and functional recovery in rat model of SCI. RSV inhibited LPS-induced inflammatory injury in PC-12 cells via increasing viability, decreasing apoptosis, and suppressing IL-1β, IL-6, and TNF-α expression. miR-132 was down-regulated after LPS treatment but up-regulated after RSV administration. miR-132 silence curbed the protective effect of RSV. The results including increase of cell growth, suppression of inflammatory response, and blocking of NF-κB and p38MAPK pathways produced by RSV were all reversed by miR-132 silence.
RSV could up-regulate miR-132 and further ameliorate inflammatory response in PC-12 cells by inhibiting NF-κB and p38MAPK pathways.
Spinal cord injury (SCI) has always been one of major medical problems due to it is really difficult to repair the central nervous system and recovery function [1]. Approximately half of SCI is induced by motor vehicle crashes and the other majorly results from falls, violence, and sports accident. SCI results in motor dysfunction, permanent paralysis and loss of sensation not only leading to serious long-term physical and psychological injuries but also causing serious social economic load [2]. The oxidative stress damage, posttraumatic inflammation, and motoneuron apoptosis and necrosis can be induced by SCI [3]. It is reported that if the SCI-induced inflammation is not suppressed, the neuronal dysfunction and cell death can be exaggerated [4, 5]. Thus, to modulate the inflammatory response is important for the treatment of SCI.
Many plant extracts have been used for medical treatment of inflammation. Resveratrol (RSV) is a natural product derived from peanuts, grapes, and many plants. The massive studies demonstrated the multiple functions and diverse activities of RSV, such as ameliorating aging-related metabolic phenotypes [6], improving insulin sensitivity and reducing oxidative stress in diabetic [7], and potently inhibiting cancer by mediating cell growth, apoptosis, angiogenesis, inflammation, and migration and metastasis by regulating multiple molecular targets [8]. More importantly, the role of RSV in SCI was investigated by a few studies, which indicated the characteristics of anti-inflammation and anti-apoptosis of RSV after SCI [9,10,11].
microRNA (miRNA), the small noncoding RNA, is regarded as a new paradigm for understanding inflammation diseases because miRNAs are important regulators in many biological processes [12, 13]. microRNA-132 (miR-132) has been shown to participate in the regulation of inflammation [14, 15]. One study explained that miR-132 suppressed lipopolysaccharide (LPS)-induced inflammation in alveolar macrophages via enhancing the cholinergic anti-inflammation pathway [14]. miR-132 was related to Toll-like receptor 4 (TLR4), regulated astrocyte activation, and modulated innate immune response in neurological diseases [16]. However, the protective effect of miR-132 on PC-12 cells has been rarely reported. The regulatory role of miR-132 in the effect of RSV was examined in this study.
Herein, we assessed the anti-inflammatory effect of RSV on the rat model of SCI or on LPS-treated PC-12 cells. The molecule mechanism of RSV in repression of inflammation response was also investigated. How miR-132 regulated the function of RSV in PC-12 cells was explored.
RSV improved inflammatory injury and functional recovery in rat model of SCI
Animal experiments were carried out to explore whether RSV exerted the protective effect on SCI. The rats were divided into four groups, including Sham group, Model group, Model+saline group and Model+RSV group. Results in Fig. 1a showed that the concentrations of IL-1β, IL-6 and TNF-α were significantly increased in SCI Model group as relative to Sham group (P < 0.001). Moreover, cell apoptosis analytical results displayed that TUNEL-positive cells and expression of pro-apoptotic regulators, p53, cleaved-Caspase-3 and Cyto. C were all promoted in Model group (P < 0.01 or P < 0.001, Fig. 1b-d). However, in Model+RSV group, we observed that the concentrations of pro-inflammatory cytokines, TUNEL-positive cells and apoptosis-associated factors were all inhibited, as compared to Model+saline group (P < 0.05 or P < 0.01, Fig. 1b-d). Additionally, evaluation of neural function showed that BBB scores in Model group were significantly declined compared to Sham group (P < 0.001). However, BBB scores in Model+RSV group were significantly increwased compared to Model+saline group (P < 0.01, Fig. 1e). More interestingly, qRT-PCR analytical results demonstrated that the expression level of miR-132 was down-regulated in Model group compared with Sham group (P < 0.05), but up-regulated in Model+RSV group compared to Model+saline group (P < 0.01, Fig. 1f). These data indicated that RSV could improve neuron protection and functional recovery in rat model of SCI. Moreover, miR-132 might be a key regulator in this process. More in vitro experiments were performed to further investigate the effect of RSV on SCI.
RSV improved inflammatory injury and functional recovery in rat model of SCI. A total of 40 SD rats were randomly divided into four groups: Sham group, Model group, Model+saline group and Model+RSV group. a The concentrations of IL-1β, IL-6, and TNF-α were increased in Model group, but decreased in Model+RSV group. b The number of TUNEL-positive cells and (c and d) the pro-apoptotic protein expressions were all enhanced in Model group, but declined in Model+RSV group. e The BBB scores were decreased in Model group, but increased in Model+RSV group. f The expression level of miR-132 was down-regulated in Model group, but up-regulated in Model+RSV group. * P < 0.05, ** P < 0.01, *** P < 0.001
RSV suppressed LPS-induced PC-12 cell injury
Cell viability was decreased by LPS treatment in a dose dependent manner (Fig. 2a). Changes of cell apoptosis were also investigated by detecting apoptotic cell rate and apoptotic factor expressions. Results showed that LPS treatment increased apoptotic cell rate (Fig. 2b) and expression levels of pro-apoptotic protein (Fig. 2c and d). Next, the effects of RSV with different concentrations on PC-12 cell viability were analyzed. Due to the viability-promoting effect of high dose of RSV (Fig. 2e), concentration of 30 μM was used for subsequent studies. Then, we analyzed the effects of RSV on LPS-treated PC-12 cells. Data demonstrated that RSV attenuated the effects of LPS on cell viability and apoptosis. Cell viability was improved in response to RSV (P < 0.05, Fig. 2f). Meanwhile, apoptotic cell rate was decreased (P < 0.01, Fig. 2g) and the expression levels of pro-apoptotic regulators, p53, cleaved-Caspase-3, and Cyto. C were all inhibited in LPS and RSV co-treated cells (Fig. 2h and i). There was no obvious change of cell apoptosis and apoptosis-associated factor in RSV alone treated cells. (Fig. 2g-i). These data suggest that RSV repressed LPS-induced loss of viability and increase of apoptosis.
RSV inhibited LPS-induced loss of viability and increase of apoptosis. LPS a decreased viability, b increased apoptotic cell rate, and (c and d) enhanced pro-apoptotic protein levels. e RSV promoted cell viability when applied higher than concentration of 40 μM. RSV with dose of 30 μM f improved cell viability, g reduced apoptotic cell rate, and (h and i) diminished pro-apoptotic protein levels. * P < 0.05, ** P < 0.01, *** P < 0.001
RSV administration inhibited LPS-induced up-regulations of pro-inflammatory factors
The inflammatory response in PC-12 cells was evaluated by detecting expressions of pro-inflammatory factors (IL-1β, IL-6, and TNF-α) after LPS or/and RSV treatment. IL-1β, IL-6, and TNF-α were all up-regulated in mRNA expression levels after LPS treatment (P < 0.01 or P < 0.001, Fig. 3a). However, all their expression levels were suppressed in RSV treated cells (P < 0.01, Fig. 3a). Similarly, the protein levels of IL-1β, IL-6, and TNF-α were all increased in LPS-treated cells, but decreased in RSV and LPS co-treated cells (P < 0.05 or P < 0.01, Fig. 3b and c). These data implied that RSV repressed LPS-induced inflammatory injury in PC-12 cells.
RSV inhibited LPS-induced overexpression of pro-inflammatory cytokines. LPS treatment induced (a) mRNA and (b and c) protein levels of IL-1β, IL-6, and TNF-α. RSV suppressed (a) mRNA and (b and c) protein levels of IL-1β, IL-6, and TNF-α. * P < 0.05, ** P < 0.01, *** P < 0.001, ns, no significant difference
miR-132 modulated the effects of RSV on PC-12 cells
For clarifying the anti-inflammatory mechanism of RSV, the role of miR-132 was analyzed. miR-132 expression was significantly declined in LPS-treated cells (P < 0.05), whereas, its expression was elevated in response to RSV treatment (P < 0.001, Fig. 4a). The modulating effects of miR-132 on growth and inflammatory reaction in PC-12 cells were detected as its expression was altered by LPS and RSV. Before this, cells were transfected with miR-132 inhibitor, and miR-132 expression was decreased after transfection (P < 0.01, Fig. 4b). According to our data, miR-132 silence impaired the protective effects of RSV on PC-12 cells. In concrete terms, significant reduction of cell viability (P < 0.01, Fig. 4c) and increase of apoptotic cell rate (P < 0.01, Fig. 4d), augmented the expression levels of pro-apoptotic regulators (P < 0.05, P < 0.01 or P < 0.001, Fig. 4e and f), as well as high levels of IL-1β, IL-6, and TNF-α (P < 0.05 or P < 0.01, Fig. 4g-i) were found in miR-132 silence group. Thus we speculated that RSV might ameliorate LPS-induced cell injury by up-regulating miR-132.
RSV ameliorated LPS injury by up-regulating miR-132. a miR-132 expression was down-regulated after LPS treatment but was increased after RSV administration. b miR-132 expression was decreased after determining miR-132-inhibitor-transfection efficiency. miR-132 knockdown c inhibited RSV-induced increase of viability, d decrease of apoptotic cell rate, as well as (e and f) suppression of pro-apoptotic protein level, and (g) mRNA/(H and I) protein levels of IL-1β, IL-6, and TNF-α. * P < 0.05, ** P < 0.01, *** P < 0.001
RSV inhibited NF-κB and p38MAPK signaling pathways by up-regulating miR-132
Whether some signaling pathways participated in the protective function of RSV was not clear. Thus we further explored this in the current study. The activations of NF-κB pathway and p38MAPK pathway were analyzed by Western blot (Fig. 5a-d). It is well-known that NF-κB and p38MAPK pathways can be activated by LPS. We next found that NF-κB nd p38MAPK pathways were suppressed by RSV because of the decreased the protein levels of p-IκBα, p-p65, and p-p38MAPK (all P < 0.01). However, the protein levels of p-IκBα, p-p65, and p-p38MAPK were all enhanced after miR-132 was knocked down (P < 0.001, P < 0.01, and P < 0.05), suggesting that NF-κB and p38MAPK pathways might be suppressed by RSV via up-regulation of miR-132.
RSV inhibited NF-κB and p38MAPK pathways. (a and b) NF-κB signaling pathway was inhibited after RSV administration but activated after miR-132 silence. (c and d) p38MAPK signaling pathway was inhibited after RSV administration but activated after miR-132 silence. * P < 0.05, ** P < 0.01, *** P < 0.001
Abundant studies focus on the therapies of SCI as SCI is a high-cost, low-curative ratio neurological disability. Inflammatory reaction incited by SCI contributes substantially to the secondary effect, with both beneficial effect and devastating effect [17]. Inhibiting harmful inflammatory response possibly contributes to protect from aggravation of the secondary injury cascade and further deterioration of neurological function [18]. Meanwhile, reducing neuronal and glial apoptosis with concomitant promotion of neurological function recovery is also associated with therapeutic targets in SCI [19]. Therefore, the aim of application of RSV in this study was to limit the inflammatory response and inhibit cell apoptosis following SCI.
Firstly, animal experiments were carried out to explore the effect of RSV on the rat model of SCI. Interesting results showed that RSV improved inflammatory injury and functional recovery in rat model of SCI. Additionally, in vitro experiment revealed that the cell injury was evaluated after LPS treatment. Cell viability and apoptosis were respectively suppressed and potentiated. The well-known apoptosis-related genes were found to be up-regulated, including p53, cleaved-Caspase-3, and Cyto. C. Moreover, the expression levels of pro-inflammatory factors were increased. However, RSV was found to reverse the negative effects mentioned above in PC-12 cells. RSV treatment produced viability-promoting and apoptosis-inhibitory effects and suppressed expressions of tested pro-inflammatory cytokines. Many investigations show that RSV possesses various biological activities, including anti-inflammatory effect [20]. RSV was showed to reduce growth inhibition, restrain apoptosis, and suppress isoflurane-induced inflammation and oxidative stress in PC-12 cells [21]. In one previous study, RSV was used to treat experimental SCI and data showed that decreased TNF-α and IL-1β expression levels, reduced TUNEL positive cells, and increased glial and motor neuron cells were found in RSV-treated group [22]. RSV potentiated the functional recovery of rat dorsal neuronal following SCI due to its activities of anti-oxidation, anti-inflammation and anti-apoptosis [9]. Therefore, all of these data were consistent with our study.
RSV displayed an important role in improving viability, reducing apoptosis, and suppressing inflammation, but how it exerted these effects was not clearly described. RSV could increase mitochondrial biogenesis and function and also protect against metabolic decline [23]. A series of miRNAs have been reported to be involved in different functions of RSV, like miR-138 [24], miR-17 [25], and miR-221 [26]. This study showed that miR-132 expression was down-regulated in LPS-stimulated cells but up-regulated after RSV treatment. miR-132 suppression impaired the effects of RSV on PC-12 cells, which indicated that the protective effect of RSV might be related to miR-132 up-regulation. Similarly, miR-132 was showed to attenuate inflammatory injury incited by LPS via targeting TRAF6 in neuron HT-22 cells [27]. miR-132 is a critical regulator mediating the integration of newborn neurons into the adult dentate gyrus. Besides, miR-132 silence in PC-12 cells induced elevated the expression levels of hundreds of genes, mostly inflammatory/immune signaling-associated genes [28]. These studies were not in conflict with our study.
Further mechanism research demonstrated that LPS-induced activations of NF-κB and p38MAPK pathways were inhibited by RSV and miR-132 up-regulation might contribute to this inhibitory effect of RSV. It is well-known that NF-κB signal pathway is an inducer of LPS as well as p38MAPK signal [29,30,31]. miR-132 attenuated LPS-induced inflammation via inhibiting the NF-κB and MEK/ERK pathways [27]. RSV suppressed the NF-κB inflammation pathway in prevention of fatty liver disease [32] and attenuated neuronal autophagy and inflammatory reaction by inhibiting the NF-κB signal in experimental traumatic brain injury [33]. RSV could also suppress the MAPK signal pathway in LPS-induced mice mastitis [34]. NF-κB p65 and p38 MAPK were key downstream factors of TLR4 and RSV was showed to block expression of TLR4 and suppress the phosphorylation of NF-κB p65 and p38 MAPK [35]. These data were consistent with our study.
Taken together, these findings demonstrated that RSV modulated miR-132 expression and attenuated LPS-induced inflammation in PC-12 cells and in rat model via promoting cell growth and suppressing expression levels of several pro-inflammatory cytokines. RSV exerted the anti-inflammation property by blocking NF-κB and p38 MAPK signaling pathways, which were also mediated by miR-132. RSV might reduce secondary damage after SCI.
Animal and experimental groups
A total of 40 SD rats (280–320 g) were obtained from Shanghai Laboratory Animal Center of Chinese Academy of Sciences (Shanghai, China). These rats were raised in cages with water ad libitum. Experimental protocols were approved by the Ethics Committee of Dalian Municipal Central Hospital. Above rats were randomly divided into four groups: Sham group, Model group, Model+saline group and Model+RSV group. The model of SCI was established according to the model of Allen's weight hit and the study from Liu et al [36, 37]. These rats were given intraperitoneal injection of trichloroacetaldehyde (300 mg/kg, i.p., Sigma-Aldrich, St. Louis, USA). Subsequently, the anesthetized rats were laid flat on the operating table, and the furs were shaved around chest and abdomen. The experiment area of rats was disinfected with alcohol, and a 3 cm incision was made at the center of the eighth thoracic spine to expose the spinal dura mater. The hitting area with the size of 3 mm × 4 mm exposed at the center of the spinal cord. After hitting, the striking instrument was quickly removed and the wound was sealed with suture. Rats in the Model+saline group received spinal cord injury and saline injection. Rats in the Model+RSV group received RSV treatment (200 mg/kg, i.p., Sigma-Aldrich) three times per day for 3 days after the injury. Rats in the Sham group received same surgical procedures but no trauma hit and RSV treatment. Finally, these rats were sacrificed by intraperitoneal injection of 100–150 mg/kg sodium pentobarbital (Sigma-Aldrich) 3 days after surgery. We tried our best to alleviate the pain of these rats.
Enzyme-linked immunosorbent (ELISA) assay
The concentrations of interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in Sham group, Model group, Model+saline group and Model+RSV group were examined by using ELISA assay based on the manufacturer instruction. In brief, the serum from the four experimental groups was collected, and the concentrations of IL-1β, IL-6 and TNF-α were measured by the corresponding ELISA kits (R&D Systems, Abingdon, UK).
TUNEL staining
TUNEL Apoptosis Detection Kit (Promega, Madison, WI, USA) was performed to detect apoptotic neurons. The number of TUNEL-positive cells was counted in five randomly chosen fields within each slide at 400× with an optic microscope (Olympus, Tokyo, Japan). The index of apoptosis was calculated according to the ratio of overall apoptotic cells and the total cells.
Evaluation of neuronal function recovery
Basso-Beattie-Bresnahan (BBB) score method was used to evaluate the functional recovery of rats in Sham group, Model group, Model+saline group and Model+RSV group. The scoring range of BBB is from zero (complete paralysis) to twenty one (normal locomotion). After 72 h post-surgery, the locomotion activity of hindlimb was evaluated.
Cell culture and treatment
PC-12 cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). They were cultured in DMEM (1 × 104 cells/ml) supplemented with fetal bovine serum (10% (v/v)), penicillin (100 U/mL) and streptomycin (100 μg/mL) at 37 °C in the incubator under suitable humidity condition. Cells were treated with LPS (1, 2, 5, and 10 μg/mL) for 12 h and RSV (ref: R5010, ≥99%, Sigma-Aldrich, St. Louis, USA) at concentrations ranging from 10 μM to 50 μM.
CCK-8 assay
CCK-8 assay was conducted to determine cell viability. Approximately 5 × 103 cells were seeded in each well of 96-well plate. After treatments, 10 μL of CCK-8 solution (Beyotime, Shanghai, China) was added to the culture medium. After further incubation for 2 h, the absorbance was measured at 450 nm using a Microplate Reader (Bio-Rad, Hercules, CA, USA).
PC-12 cells were gathered by centrifugation (2000 g for 10 min) and then they were rinsed with PBS. Afterwards, PC-12 cells were suspended in 300 μL binding buffer followed by adding 5 μL Annexin V-FITC solution. Then cells were incubated with Annexin V-FITC at room temperature in the dark for 15 min. After incubation, 5 μL PI solution was added in the cell suspension. Finally, 200 μL binding buffer was supplemented and flow cytometry analysis was conducted immediately. FACScan (Beckman Coulter, Fullerton, CA, USA) was used and the data were analyzed by using FlowJo software.
miRNA transfection
For miR-132 silence transfection, miR-132 inhibitor (5'-AGU AAC AAU CGA AAG CCA CGG U-3′) synthesized by GenePharma Co (Shanghai, China) was transfected into cells using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA) in this study. Transfected cells were harvested after post-transfection of 48 h.
qRT-PCR analysis
The qRT-PCR for miR-132 was performed in a CFX96 Real-Time PCR Detection System (Bio-Rad) as described previously [38]. The primer sequences for miR-132 were followed as: forward, 5'-ACC GTG GCT TTC GAT TGT TA-3′; reverse, 5'-GGC GAC CAT GGC TGT AGA CT-3′. Total miRNAs in treated or transfected cells was extracted by using miRNeasy Mini Kit (Qiagen, Shenzhen, China). RNAs were reverse transcribed with PrimeScript Reverse Transcriptase (Takara, Dalian, China). For mRNA levels of IL-1β, IL-6, and TNF-α, total RNAs were extracted by using Trizol reagent and treated with DNAse I (Promega, Madison, WI, USA), and were reverse transcribed in a reaction system containing random primers and M-MLV reverse transcriptase. Subsequently, the cDNAs were amplified by using RT-PCR with SYBR green Master Mix. U6 was used as the internal control for miR-132 expression analysis and β-actin was used as the internal control for determining expression of IL-1β, IL-6, and TNF-α. Their expression levels were calculated by relative quantification 2−△△CT method.
Total protein was extracted from PC-12 cells; and after quantification, 50 μg protein was separated by 12% SDS-PAGE and transferred to the PVDF membrane. The membranes were incubated with appropriate primary antibodies prepared in 5% blocking buffer at a dilution of 1:1000 at 4 °C overnight. The primary antibodies used in this study were displayed as follows: p53 (ab26), pro-Caspase-3 (ab44976), cleaved-Casapse-3 (ab13847), Cyto. C (ab133504), IL-1β (ab9722), IL-6 (ab9324), TNF-α (ab6671), IκBα (ab32518), p-IκBα (S36, ab133462), p65 (ab16502), p-p65 (S536, ab86299), p38MAPK (ab31828), p-p38MAPK (T180, ab178867), and β-actin (ab8226, Abcam, Cambridge, MA, USA). After incubation, the PVDF membranes were further incubated with the horseradish peroxidase-conjugated second antibody for 1 h at room temperature. Subsequently, signals were captured by using the enhanced chemiluminescence reagent of Lumi-Light Western Blotting Substrate (Sigma-Aldrich, St, Louis, MO, USA), and were scanned by using the UMAX Vista S6E Flatbed Scanner (UMAX data systems inc., Hsinchu, Taiwan). The intensity of the target band was analyzed by Image Lab™ Software (Bio-Rad, Hercules, CA, USA).
Data were expressed as the mean ± SD of at least three independent experiments. We evaluated the data with a one-way analysis of variance (ANOVA) followed by Sidak's post hoc test for multiple comparisons. The data with P value less than 0.05 was considered significant.
LPS:
Lipopolysaccharide
miR-132:
microRNA-132
RSV:
SCI:
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Wang HD, Shi YM, Li L, Guo JD, Zhang YP, Hou SX. Treatment with resveratrol attenuates sublesional bone loss in spinal cord-injured rats. Br J Pharmacol. 2013;170(4):796–806.
Dai R, Ahmed SA. MicroRNA, a new paradigm for understanding immunoregulation, inflammation, and autoimmune diseases. Translational research : the journal of laboratory and clinical medicine. 2011;157(4):163–79.
Singh RP, Massachi I, Manickavel S, Singh S, Rao NP, Hasan S, Mc Curdy DK, Sharma S, Wong D, Hahn BH, et al. The role of miRNA in inflammation and autoimmunity. Autoimmun Rev. 2013;12(12):1160–5.
Liu F, Li Y, Jiang R, Nie C, Zeng Z, Zhao N, Huang C, Shao Q, Ding C, Qing C, et al. miR-132 inhibits lipopolysaccharide-induced inflammation in alveolar macrophages by the cholinergic anti-inflammatory pathway. Exp Lung Res. 2015;41(5):261–9.
Shaked I, Meerson A, Wolf Y, Avni R, Greenberg D, Gilboa-Geffen A, Soreq H. MicroRNA-132 potentiates cholinergic anti-inflammatory signaling by targeting acetylcholinesterase. Immunity. 2009;31(6):965–73.
Kong H, Yin F, He F, Omran A, Li L, Wu T, Wang Y, Peng J. The effect of miR-132, miR-146a, and miR-155 on MRP8/TLR4-induced astrocyte-related inflammation. Journal of molecular neuroscience : MN. 2015;57(1):28–37.
Hausmann ON. Post-traumatic inflammation following spinal cord injury. Spinal Cord. 2003;41(7):369–78.
Cronin M, Anderson PN, Cook JE, Green CR, Becker DL. Blocking connexin43 expression reduces inflammation and improves functional recovery after spinal cord injury. Mol Cell Neurosci. 2008;39(2):152–60.
Ferrer I, Barrachina M, Tolnay M, Rey MJ, Vidal N, Carmona M, Blanco R, Puig B: Phosphorylated protein kinases associated with neuronal and glial tau deposits in argyrophilic grain disease. Brain pathol. 2003, 13(1):62–78.
Inoue H, Nakata R. Resveratrol targets in inflammation. Endocr Metab Immune Disord Drug Targets. 2015;15(3):186–95.
Hu W, Yang E, Ye J, Han W, Du ZL. Resveratrol protects neuronal cells from isoflurane-induced inflammation and oxidative stress-associated death by attenuating apoptosis via Akt/p38 MAPK signaling. Experimental and therapeutic medicine. 2018;15(2):1568–73.
Senturk S, Yaman ME, Aydin HE, Guney G, Bozkurt I, Paksoy K, Abdioglu AA. Effects of Resveratrol on Inflammation and Apoptosis after Experimental Spinal Cord Injury. Turkish neurosurgery. 2017.
Price Nathan L, Gomes Ana P, Ling Alvin JY, Duarte Filipe V, Martin-Montalvo A, North Brian J, Agarwal B, Ye L, Ramadori G, Teodoro Joao S, et al. SIRT1 is required for AMPK activation and the beneficial effects of resveratrol on mitochondrial function. Cell Metab. 2012;15(5):675–90.
Zhang Y, Du X, Li W, Sang H, Qian A, Sun L, Li X, Li C. Resveratrol improves endothelial progenitor cell function through miR-138 by targeting focal adhesion kinase (FAK) and promotes Thrombus resolution in vivo. Medical science monitor : international medical journal of experimental and clinical research. 2018;24:951–60.
Pan J, Shen J, Si W, Du C, Chen D, Xu L, Yao M, Fu P, Fan W. Resveratrol promotes MICA/B expression and natural killer cell lysis of breast cancer cells by suppressing c-Myc/miR-17 pathway. Oncotarget. 2017;8(39):65743–58.
Wu F, Cui L. Resveratrol suppresses melanoma by inhibiting NF-kappaB/miR-221 and inducing TFG expression. Arch Dermatol Res. 2017;309(10):823–31.
Ji YF, Wang D, Liu YR, Ma XR, Lu H, Zhang BA. MicroRNA-132 attenuates LPS-induced inflammatory injury by targeting TRAF6 in neuronal cell line HT-22. J Cell Biochem. 2018.
Luikart BW, Bensen AL, Washburn EK, Perederiy JV, Su KG, Li Y, Kernie SG, Parada LF, Westbrook GL. miR-132 mediates the integration of newborn neurons into the adult dentate gyrus. PLoS One. 2011;6(5):e19077.
Lawrence T. The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harb Perspect Biol. 2009;1(6):a001651.
Ghosh S, Hayden MS. New regulators of NF-κB in inflammation. Nat Rev Immunol. 2008;8:837.
Bode JG, Ehlting C, Häussinger D. The macrophage response towards LPS and its control through the p38MAPK–STAT3 axis. Cell Signal. 2012;24(6):1185–94.
Tian Y, Ma J, Wang W, Zhang L, Xu J, Wang K, Li D. Resveratrol supplement inhibited the NF-kappaB inflammation pathway through activating AMPKalpha-SIRT1 pathway in mice with fatty liver. Mol Cell Biochem. 2016;422(1–2):75–84.
Feng Y, Cui Y, Gao JL, Li MH, Li R, Jiang XH, Tian YX, Wang KJ, Cui CM, Cui JZ. Resveratrol attenuates neuronal autophagy and inflammatory injury by inhibiting the TLR4/NF-kappaB signaling pathway in experimental traumatic brain injury. Int J Mol Med. 2016;37(4):921–30.
Zhang X, Wang Y, Xiao C, Wei Z, Wang J, Yang Z, Fu Y. Resveratrol inhibits LPS-induced mice mastitis through attenuating the MAPK and NF-κB signaling pathway. Microb Pathog. 2017;107:462–7.
Chin YT, Cheng GY, Shih YJ, Lin CY, Lin SJ, Lai HY, Whang-Peng J, Chiu HC, Lee SY, Fu E, et al. Therapeutic applications of resveratrol and its derivatives on periodontitis. Ann N Y Acad Sci. 2017;1:101–8.
Wrathall JR, Pettegrew RK, Harvey F. Spinal cord contusion in the rat: production of graded, reproducible, injury groups. Exp Neurol. 1985;88(1):108–22.
Liu J, Yi L, Xiang Z, Zhong J, Zhang H, Sun T: Resveratrol attenuates spinal cord injury-induced inflammatory damage in rat lungs. Int J Clin Exp Pathol. 2015;8(2):1237–46.
Zhao Rb, Zhu Lh, Shu JP, Qiao LX, Xia ZK. GAS5 silencing protects against hypoxia/ischemia-induced neonatal brain injury. Biochem Biophys Res Commun. 2018;497(1):285–91.
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Department of Spinal Surgery, Dalian Municipal Central Hospital, No.826, Xinan Road, Shahekou District, Dalian, 116033, Liaoning, China
Guiqi Zhang
, Yi Liu
, Lichen Xu
, Chunhe Sha
, Haibin Zhang
& Weibing Xu
Search for Guiqi Zhang in:
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GZ, YL, LX, CS and HZ carried out the experiments. GZ drafted the paper, YL revised the paper. WX participated in the coordination and guidance. All authors have read and approved the final manuscript.
Correspondence to Weibing Xu.
Zhang, G., Liu, Y., Xu, L. et al. Resveratrol alleviates lipopolysaccharide-induced inflammation in PC-12 cells and in rat model. BMC Biotechnol 19, 10 (2019) doi:10.1186/s12896-019-0502-1
p38MAPK
Cellular and tissue engineering

redpajama_set_name: RedPajamaCommonCrawl


text:
Q: padding list of tensors with varied size
I have a Python list of 1-D tensors with different sizes. For example,
[ tf.constant([1]), tf.constant([2,3]), tf.zeros([4,4,4]) ]

is there a way to pad the tensors so they have the same size, like the following,
[ tf.contant([1,0,0]), tf.constant([2,3,0]), tf.constant(4,4,4) ]

then I can use tf.stack to turn them back into one tensor. These tensors are in the middle of a computation graph: they are NOT from the input.

redpajama_set_name: RedPajamaStackExchange


text:
Table of Contents » Title 19.2. Criminal Procedure » Chapter 13. Grand Juries » Article 3. Special Grand Juries » § 19.2-213. Report by special grand jury; return of true bill
§ 19.2-213. Report by special grand jury; return of true bill.
At the conclusion of its investigation and deliberation, a special grand jury impanelled by the court on its own motion or on recommendation of a regular grand jury shall file a report of its findings with the court, including therein any recommendations that it may deem appropriate, after which it shall be discharged. Such report shall be sealed and not open to public inspection, other than by order of the court.
A majority, but not less than five, of the members of a special grand jury convened upon request of the attorney for the Commonwealth must concur in order to return a "true bill" of indictment. A "true bill" may be returned upon the testimony of, or evidence produced by, any witness who was called by the grand jury, upon evidence presented or sent to it.
1975, c. 495; 1978, c. 638; 2001, c. 4.


text:
Criminal Justice Department celebrates 40 years
October 27, 2014 12:06 am by Eavn Hale
With faculty, administration, students and alumni, members of the Criminal Justice Department, both present and past, celebrated 40 years of excellence at Grambling State University during Grambling State University's Homecoming activities.
The event held in the Betty E. Smith Nursing Auditorium on Friday, Oct. 17, was a Homecoming reunion and fundraiser where alumni returned to their alma mater to mingle with students, share success stories and celebrate four decades worth of service at GSU. Items such as autographed footballs, portraits of Eddie G. Robinson and gift baskets were just a few items raffled in a silent auction. The department was able to raise over $15000, which will go towards criminal justice majors and organizations such as Alpha Phi Sigma Inc.
"I thought it was an awesome program," said Mae Conley, a 28-year criminal justice professor at GSU. "Our students were exposed to what they can accomplish if they work hard in the classroom," said Conley, a graduate of Mississippi Valley State. "I was so proud when I saw all my prior students come back and flash their success with the present students."
On the program were several speakers whom since attended Grambling and are now prominent figures in the criminal justice field. One of the speakers, Nicole Sinegar is one out of 147 African American woman in FBI, which has only 500 African Americans altogether. In total, there are 13000 agents and that is just one of the accomplishments achieved by the alumni. Each speaker's message was different but the topic, nevertheless, remained the same sharing words of encouragement and valuable info about securing a job with the federal government.
"It's not about what you know, it's about who you know," said Keith Brach, a GSU alumni whom is now the assistant director of residential services at Esquire in Houston. "Network, dare to be different."
"Success is a formula, not a fantasy," said Reller Jones, a former criminal justice professor and now campus volunteer at GSU.
The alumni speakers wanted students to be proactive in their studies as well as utilize their time in college wisely. They shared with the students their experiences at GSU as well as knowledge about the different criminal justice fields.
"Listening to the alumni, I learned about some of the requirements I need to be successful," said Deandrea Federick, a sophomore criminal justice major from Bossier City.
Although each speaker's message lamented similar meaning, what each student took away from it differed.
"I learned that when you go in for an interview, you are interviewing before you even open your mouth and speak to someone," said Amber Williams, a senior criminal justice major from Dallas. "It's about who you know to get you in the door and what you know to keep you there."
The alumni that came back to Grambling to celebrate this event represented government agencies well because there were so many different branches from state troopers to special agents to border patrol and so on. Not only did the alumni donate time but also they each donated an allocated cash amount back to the department.
"It just goes to show the foundation of Grambling State University is always willing to help each other," said Conley about the success of the event. "As proud as I was, it was a humbling experience because all of our students came back and they didn't have to. With all of their success they still wanted to give back to the students."
Tags: Criminal justice


text:
A Cheaper Galaxy Note smartphone might be on the way
Samsung Android
Published: Sep 25, 2019, 7:39 AM
Joshua Swingle
The Galaxy Note 10 series is far from what you'd consider affordable and the S Pen, in turn, is far from what within reach. But come next year, this entire situation could change.
The phone should be cheaper than the Galaxy Note 10
According to SamMobile, Samsung is developing a device with the model number SM-N770F that doesn't seem to fit into any of the brand's existing lineups. The SM-Nxxx range is typically attached to Galaxy Note-branded models yet the company's most recent devices carry the SM-N9xx numbers. This leads SamMobile to believe that what Samsung is developing at the moment may be a cheaper offering.
For reference, the soon-to-be-released Galaxy Fold carries the model number SM-F900F. However, the South Korean giant is understood to be working on a more affordable version known internally as SM-F700F that'll arrive next year. This model number strategy, as you may have noticed, coincides perfectly with the SM-N770F number associated with the upcoming S Pen-bearing phone.
Don't expect a $1,000 price tag
Unfortunately for us, very little is known about the device right now because development appears to be in the early stages. However, SamMobile did uncover some details which suggest it includes 128GB of internal storage as standard. Whether this is expandable or not, however, will remain to be seen.
What kind of phone could this be?
When it comes to determining what kind of device this will be, it really is a guessing game at this stage. Nevertheless, there do appear to be a couple possibilities.
A Galaxy Note 10e-like device
Rumors suggest Samsung is going to merge the Galaxy S and Galaxy Note lineups next year. The combined series will apparently be called Galaxy One and it may include a Galaxy Note 10e-like device. This could be positioned below the Galaxy S11 but above the Galaxy S11e and would likely represent an S Pen-bearing version of the latter. To help make it a more attractive purchase, some other extras and upgrades could be included.
If this is the case, the smartphone will likely debut in late February alongside the rest of the Galaxy S11 series. An announcement date is yet to be confirmed but Samsung's usual timeline suggests the Unpacked event may take place on Wednesday, February 19. The device may be available to pre-order on Friday, February 21 and begin shipping a couple of weeks later on Friday, March 6.
A Galaxy S10e-like phone but with S Pen capabilities?
Pricing obviously isn't yet confirmed but some educated guesses can still be made. Assuming the Galaxy S11e retails at $699 like the iPhone 11, the Galaxy Note 10e could slot in at $799. This would then allow Samsung to position the Galaxy S11 at $899 and the Galaxy S11+ at $999.
A Mid-range Galaxy Note phone
The S Pen is often the sole reason consumers purchase Galaxy Note flagships over Galaxy S models. However, not everyone wants to spend $1,000 or more on a smartphone. By introducing a model that's positioned below the likes of the Galaxy S11e, Samsung would be opening up the S Pen to an entirely new crowd. This, in turn, could help drive sales and better differentiate the South Korean giant's models from those of Huawei and Xiaomi.
Unfortunately, the unveiling date in this scenario remains a total mystery because Samsung could either announce it at MWC 2020 in February or at any other point in the following months. The South Korean company may also skip the pre-order period due to the nature of the phone and simply start shipping it on the announced date.
Samsung's Galaxy S and Galaxy Note lines could merge next year
Sep 15, 2019, 3:57 PM, by Joshua Swingle


Meyer, V. (ed.), Frontiers in Offshore Geotechnics III: proceedings of the Third International Symposium on Frontiers in Offshore Geotechnics (ISFOG 2015), Oslo, Norway, 10-12 June 2015, CRC Press, Boca Raton, pp. 337-342.
This paper presents resistance envelope equations for open-ended caissons, considering failure of the soil inside the caisson (i.e internal scooping). Equations for assessment of failure envelopes of caissons installed in undrained soil were developed by Kay&Palix (2011) for various embedded Length (L) over Diameter (D) ratios and varying undrained shear strength profiles. One of the assumptions in Kay & Palix (2011) is a solid caisson, which implies no possibility of internal scooping failure. 3D finite element software was used to assess equations for VHM failure envelopes for open-ended caissons with various embedment (L/D) ratios in undrained soil with varying undrained shear strength profiles. The assessments show a significant reduction in VHM failure envelopes for caissons with L/D ratios less than 2, that are installed in non-uniform soil conditions. The equations can be used for design purposes.


UNITED STATES—The House Judiciary Committee met in Washington D.C. on Tuesday, April 9 to discuss the increase in hate crimes in the country. House Judiciary Committee Chairman, Jerrold Nadler gave opening remarks.
I said it yesterday, and I meant it: This woman, Candace Owens, is the TRUTH! And that's why Democrats and their buddies in the propaganda media are jumping through hoops to smear her.
Owens said she was a victim of a hate crime as a teenager. An ANTIFA group decided she was not fit to eat in the same restaurant they were eating in. She brought her grandfather, Robert Owens who grew up as a sharecropper who lived through the Ku Klux Klan shooting bullets through the windows of his childhood home.
"There isn't a single adult today that in good conscience [that] would make the argument that America is a more racist, white nationalist society than when my grandfather was growing up and yet we are hearing these terms center around today because what they want to say is that brown people need to be scared which seems to be the narrative that we hear every four years right ahead of a Presidential election," said Owens.
Congressman Ted Lieu shared a partial recording of a comment Owens previously about Hitler. Lieu referred to a statement Owens made in a speech explaining globalism, and not wanting globalism in 2018.
"When we say nationalism, the first thing people think about, at least in America, is Hitler. He was a national socialist. If Hitler just wanted to make Germany great and have things run well, Okay fine. The problem is that he had dreams outside of Germany," said Owens in the recording.


text: Q: Pow.cx with xip.io Rails app still uses .dev for CSS & JS assets I've successfully accessed a local Rails app, setup using pow.cx, via a mobile device and Edge Inspect using xip.io. Great. Only problem, all my CSS & JS assets still use the .dev domain, rather than the xip.io domain, so the pages appear completely un-styled, with no javascript.


text:
2. NEA Seminar (1)
Nuclear Energy Agency of the OECD (NEA) (1)
BIOLOGICAL EFFECTS (1)
BIOLOGICAL RADIATION EFFECTS (1)
The ICRP radiation protection philosophy and its applications
Beninson, D.
The Interface in Nuclear Safety and Public Health1991
[en] The ICRP philosophy of radiation protection is described in the main Recommendations issued periodically. The paper reviews some issues of this philosophy on the bases of the new Recommendations expected to be approved by the end of the year. In these Recommendations, the Commission will not change the basic principles of its system of radiation protection, namely that practices causing exposures should be justified, the protection arrangements should be optimized, and the individual exposures should be restricted by dose limits, source-related dose constraints or prescribed limits, as appropriate. As an important conceptual development, the Commission will clearly differentiate in its recommendations between practices giving rise to radiation exposures, calling for the application of the system of protection, and situations where existing radiation exposures require decisions on remedial actions. In its review of radiobiology and epidemiology, the Commission concludes that new data and new interpretation of earlier information now indicate with reasonable certainty that the risks associated with ionizing radiation are about three times higher than they were estimated to be a decade ago. This increase calls for some quantitative changes in the Commission's recommendations. One such change to be recommended by the Commission is a reduction of the dose limit for occupational exposure. The current figure of 50 millisievert in a year will be reduced to 20 millisievert in a year, with some provisions to allow year-to-year flexibility. The current limit for the long-term dose for the public (1 millisievert per year averaged over a lifetime) will be retained, but increased protection will be provided by limiting the averaging period to five years and recommending the use of additional source-related constraints
RADIATION PROTECTION AND DOSIMETRY (C5300)
Nuclear Energy Agency, 75 - Paris (France); 249 p; ISBN 92-64-03349-1; ; 1991; p. 64-75; Organisation for Economic Co-operation and Development; Paris (France); 2. NEA Seminar; Paris (France); 12-13 Sep 1990
Nuclear Energy Agency of the OECD (NEA)
BIOLOGICAL RADIATION EFFECTS, DOSE EQUIVALENTS, DOSE LIMITS, DOSE-RESPONSE RELATIONSHIPS, EPIDEMIOLOGY, HUMAN POPULATIONS, ICRP, NEOPLASMS, OCCUPATIONAL EXPOSURE, OPTIMIZATION, PERSONNEL, PROBABILISTIC ESTIMATION, RADIATION DOSES, RADIATION PROTECTION, RECOMMENDATIONS, RISK ASSESSMENT
BIOLOGICAL EFFECTS, DISEASES, INTERNATIONAL ORGANIZATIONS, POPULATIONS, RADIATION EFFECTS, SAFETY STANDARDS, STANDARDS


text:
Path Puzzles
Check out PathPuzzles.com
Get the App from iTunes
Path Puzzles is a game built for iOS that challenges players to draw a path across a puzzle grid from one entrance to another. Game play and rules were created by Rod Kimball, first as a paper and pencil logic puzzle. Working with Kimball, Three Byte Intermedia was excited to build a phone app that would allow more people to play these puzzles.
Each puzzle is a different grid pattern, with numbers placed adjacent to playable squares. To beat a puzzle, the player must draw a red path from one opening (entrance) to another while satisfying the restrictions imposed by the numbers. A valid path must run through the exact number of cells in the row and column adjacent to the number, while never going through a cell twice.
Rod Kimball discusses his experience working on the app:
My goal with the Path Puzzles app was to make a marketing tool for the Path Puzzles book. To this end I wanted to create an interface that would allow solvers to have the same experience of using logical deduction as they would with pencil and paper. In addition, players would benefit from the technical and practical advantages of a touch interface. In working out the nuts and bolts, the Three Byte guys are wonderful to work with. Every idea I had was fully explored. They introduced many great ideas as well, but I always felt in control of the final product. Throughout the process, Three Byte has done everything possible to make sure the Path Puzzles app is as successful as it can be. It felt great to be able to place the technical burden completely in somebody else's hands, freeing me to simply brainstorm and create. The Three Byte team took on that burden gladly.
To make sure both leading mobile platforms were supported, Three Byte developed this game on top of the PhoneGap cross platform framework (powered by Apache's Cordova). PhoneGap allows developers to build web applications, while rendering them on mobile device as native applications.
Different mobile platforms use different native features, so Path Puzzles cannot run exactly the same on different operating systems. Kimball wanted the app to be free to download, so players could start with the base level set, before purchasing additional levels. To do this, Path Puzzles has to integrate with both the iTunes App Store to allow gamers to make in-app purchases to buy more puzzles. Each app market works differently, with different rules and APIs. In addition, different iPhones and iPads, each with individual screen dimensions, must be supported. CSS3 technology makes it easier than ever to support a wide array of different display shapes and sizes.
To provide a puzzle builder with an easy way to build new puzzles which can be included in add-on level packages, Three Byte developed a web based board builder application. This application gives level designers all the tools they need layout new puzzles, play them in a sample game environment, while testing the validity and beatability of boards algorithmically on Three Byte's server.
Here's Rod introducing Path Puzzles on YouTube:


Castle Tree & Landscaping can look after and maintain your private gardens or commercial business property on a regular basis. We work with Residential and Commercial properties in the Northamptonshire area. We offer a wide range of skills to complement your private garden or grounds ensuring they are left clean, tidy and fresh. We typically visit your garden or commercial property every two weeks, enabling us to keep a close eye on all the needed aspects of your maintenance service. However, we can also work on a more suitable schedule, tailored to your requirements.
We will ensure your lawn is kept in immaculate condition all year round. Regular cutting, edge trimming and weeding ensure a lush green garden.
Our expert employees trim and prune hedges, bushes and plants at the correct time of year. This ensures a healthy and tidy garden throughout each season even during winter.
Not only do we clear all garden waste we can also clean your patios, driveways and decking areas. Your outside areas will be left clean, tidy and maintenance free all year round.


There is a very famous saying that the beauty is fade but the knowledge you have is for your entire life and we can say knowledge is forever, that is why according to me knowledge is very important and better as compare to beauty, Because knowledge ultimately gives beauty in personality.knowledge has a great power. One can communicate on the basis of his knowledge not the behalf of his/her beauty. Knowledge simply makes your personality more dynamic, energetic and beautiful but beauty can not makes you a good knowledgble person. so if you have a very good amount of knowledge then people will definetly do your respect and listen what ever you say with full concentration and intrest. And it is proved that if you have knowledge then you can talk without hesitation and you can pass your comment about what you know and can tell others that what is right and what is wrong, and definetly people will listen to you carefuly and learn from you because they know that you are a person who have some sort of knowledge. Knowledge gives you the power to influnce others and it gives you the power to stand in a big crowd with the high level of confident. If anybody has knwoedge then people will see him/her from positive point of view. As i mention earlier that through knowledge you can influnce others, so we can say that through knowledge you can be a good leader, because the big and chalenging role of leader is to influnce others and without good knowledge it is imposible to influnce people.
Here comes the main question if knowledge is soo much important then from where to get knowledge, actually knowledge comes from lots of different areas, now its depend on people that from which area they want to get knowledge, like some people believe on research so they are getting knowledge from different resarches, some believe on learning by doing things and get knowledge, and students work hard to get knowledge and for that they read lots of different informative books and knwoedge based articles. We need knowledge in the every prespect of life if you want a good job then for that you must need good knowledge regarding that particular job, can you get a good job on the basis of your beauty? Sounds like stupid, definetly No. Thats why i say that knowledge is very much important then beauty.
In some cases beauty has their certain importance, but knowledge let you shine like a star in any where any place of the world, Actually knowledge is like a candle which gives you light in the dark and gives you brightness in your life.
Through knowledge you can take right decision at right time, and because you take decision on the basis of your knowledge wether it is the knowledge of books or the knowledge that you get from your exprience, you stand like a strong tree on your decision because you know there is less risk of loss because of your knowledge regarding the decision you take, and definetly you will be very much confident on your decision because of your knowledge.
In last i want to say that knowledge makes you stand on the most higher position, remember one thing in your life that, walking in the sunlight can destroy your beauty but there is noting in this world that can destroy your knowledge. So always try to get maximum amount of knowledge wether it is related to your academics or not, but remember one thing always utilize your knowledge in the proper way, and try to give to your society as much as you can.


text:
Articles from the August 4, 2022 edition
By Katelin Carroll News August 4, 2022
Cops and Kids Whiffle Ball Match
On Friday, July 22, 2022, Hope Hohenwald hosted their second annual Cops and Kids Whiffle Ball Match where kids play a game of whiffle ball against HPD Officers, Sheriff Deputies and even a Tennessee...
Commission sets Property Tax Rate
The County Commission met on Monday, July 25, 2022, to discuss quarterly reports from other county departments and budgets/appropriations for the upcoming fiscal year. During the alloted time for public input, Commissioner Brian Peery called upon...
News August 4, 2022
TBI investigates death of Lewis County inmate
The Tennessee Bureau of Investigation officially announced that they have launched an investigation into the death of 23-year-old Donavan Conner. "We can confirm TBI special agents are investigating the death of Donavan Connor (DOB 4/26/1999),...
Postal Service To Feature Elephants on Forever Stamps
The Elephant Sanctuary in Tennessee is excited to announce that the United States Postal Service's First Day of Issue Dedication Ceremony for the new Forever Stamp titled Elephants will take place at...
Election Day is Thursday, August 4, 2022. Go out and vote! Polls open at 9:00 a.m. and close at 7:00 p.m....
Lewis County Schools Media Day fundraiser
Lewis County Schools will hold a Media Day fundraiser on August 6, 2022 at 11:00 a.m. LCMS and LCHS football players, cheerleaders, and band members will be announced at the high school. There will be a dunking booth, Panther gear, BBQ plates, yard...
Obituaries August 4, 2022
Donavan Conner
April 26, 1999 ~ July 26, 2022 On July 26, 2022, Donavan Nathaniel Blake Conner left this earthly world at the age of 23. Donavan was born in Maury County on April 26, 1999, to Terri Adkins and the la...
Winford Leon Self
June 28, 1951 ~ July 29, 2022 Winford Leon Self, age 71, of Summertown, passed away Friday, July 29, 2022, at his residence. He was a native of Lawrence County, retired from Cytec Industries, and of...
Jerry Don Ammons
July 27, 1940 - July 25, 2022 Jerry Ammons, of Hohenwald, passed away on Monday, July 25, 2022, two days shy of his 82nd birthday. He was born in Hohenwald on July 27, 1940 to the late Carlos Don...
Flossie Bell Walker
February 2, 1939 ~ July 30, 2022 Flossie Bell Walker, age 83, of Summertown, passed away Saturday, July 30, 2022, at Maury Regional Medical Center in Columbia. She was a native of Summertown, a homemaker, and a member of Napier Pentecostal Church....
Danny Neal Bates
July 16, 1941 ~ July 30, 2022 Danny Neal Bates, 81, passed away Saturday morning July 30, 2022, at Lifecare in Centerville. Danny was born July 16, 1941, to the late Jason Clifford and Pauline Hensley Bates of Hohenwald, the first son and second...
Betty Lee Jones
August 25, 1940 - July 28, 2022 Betty Lee (Davis) Jones, a life-long resident of the Oak Grove Community in Lewis County, passed peacefully from this life to heaven on Thursday, July 29, 2022, at the...
Anchor Club donations
Members of the Anchor Club recently held a car wash. Proceeds went to Lewis County Library expansion fund and school supplies for needy students at LCHS. Anchor club is a service organization for...
Main Street Fabrics open for business with new owners
New business owners Dawn and Eric Talley have now reopened Main Street Fabrics! Dawn hosted the soft opening alongside her family on Friday, July 29, 2022, where they offered great deals and steals...
Business August 4, 2022
Public Notice; Beer Board Meeting
The Lewis County Beer Board will convene on Tuesday, August 16, 2022 at the Lewis County Courthouse in the Courtroom at 4:30 p.m. To be reviewed is the application for on-premises consumption filed by George A. Barber of the Lewis County Elks Lodge...
Descendants of the Patriot John Talley gather at Lomax Church of Christ
Descendants of John and Jane Talley held a reunion on July 23, 2022, at the Lomax Church of Christ, Service Center, in Hohenwald. The theme of the event was "deep roots, many branches", a testament...
By Dorothy Warf News August 4, 2022
Gordonsburg/ Little Swan News
We have had a wet week and that is good although there were lots of thunderstorms around the area but no damage that I know of except lots of twigs and limbs around the area. We are thankful that we did not have the flooding that Kentucky is facing...
Class of 1957 celebrate 65th year anniversary
The Lewis County High School Class of 1957 met at the Freedom Holy Temple Church to celebrate the 65th anniversary of their graduation on July 23, 2022. The class members started arriving about 11...
A hole in...
Timmie Hinson managed a Double Eagle on hole five while golfing with his friends David Barber, Kenny Collier and CW Carroll at the Hohenwald Golf Course on Tuesday, July 25, 2022. If you or a friend has earned a hole-in-one or another great score...
Freed-Hardeman University's Horizons Camp strengthens young Christian leaders
Freed-Hardeman University students spent a week mentoring young Christian leaders during Horizons this month on FHU's campus. Each summer more than 1,000 campers, 7th through 12th graders, attend Horizons to connect with fellow Christians, learn...
By Evelyn 'E.T.' Trevathan News August 4, 2022
Well it is a beautiful morning here in our little valley, I have got my coffee and I am on the front porch swing. The cats are finished with breakfast and one of them has decided to sit in the swing with me, she is the one that was really sick but...
By Callie Dunn Calendar August 4, 2022
Celebrating...
Birthdays Teresa Perry, August 4 Kendy Roden Lindley, August 4 Mary Ellean Stephens, August 4 Emma Grace Hensley, August 4 Paulette Rone, August 5 Karen Pipkins Carroll, August 5 Lisa Ammons Chambers, August 5 Donnie Breece, August 5 Candace...
By Crystal Nash News August 4, 2022
Glimpses of Hohenwald's Past
Banner dedicated to elected mayors and councilmen
This banner is dedicated to elected mayors, councilmen past, present and future of Hohenwald, Tennessee, in loving memory of Mayor S. Dol Willis, my Dad. I grew up with a father that loved Hohenwald...
Lewis County Public Library and Archives staff earns new certification
The Lewis County Public Library and Archives is pleased to announce all staff members have earned the Tennessee State Library and Archives' Core Competencies certification. Lewis County library staff...


Hidden Veggie Quesadilla–A cheese quesadilla is given a healthy make-over with the addition of a layer of butternut squash puree. This Healthy Quesadilla is Served with Cilantro Lime Cream and is the perfect way for picky eaters to enjoy veggies.
My daughter could live on cheese if we let her.
She would live on Grilled Cheese or Mac and Cheese if we let her.
She sort of sounds like a typical young child, doesn't she?
Most toddlers and elementary aged children have a love affair with a few simple foods and it can be a daunting task to include new things or vegetables. Maybe you can relate?
And while I STRONGLY suggest getting your kids into the kitchen with you to cook and to invite them to try new things daily, I am NOT opposed to sneaking in a bit of healthy where I can, like Mac and Cheese Muffins or Veggie Quesadillas.
This is not your average cheese quesadilla.
A little bit of veggies are tucked inside a typical cheese quesadilla, to make it a bit healthier.
I call these healthy quesadillas, "Hidden" Veggie Quesadillas, but it truth, my kids KNOW there are veggies there.
I believe it is important to let kids know that veggies can taste good and SHOULD be included in their diets.
But these cheese quesadillas are a great way to "sneak" in some veggies without the flavor of the quesadilla being overwhelmed with the taste of vegetables.
Start with a simple cheese quesadilla, which is typically a sure-fire win for most kids, and add in a layer of butternut squash puree.
Spread one tortilla (corn, flour, or gluten-free) with a layer of butternut squash puree.
Cook quesadilla on griddle or in microwave just long enough to melt the cheese.
I find that butternut squash is one of the best vegetables to pair with a cheese quesadilla. The butternut squash puree is the same color as the cheddar cheese, so it is easily masked in appearance. Also, butternut squash has such a mild flavor, that it is easily hidden from young palates as well.
The easiest way to roast butternut squash is to roast BEFORE cutting.
Place the WHOLE butternut squash in the oven and bake for 1 hour at 350 degrees.
Remove it and slice the squash open–it is SO soft, it almost cuts with a butter knife.
Scoop out the seeds, and place flesh of squash in large bowl.
You can also place in blender of food processor, but not a necessary step at this point.
Freeze butternut squash puree in small portions for later use.
*Feel free to use pumpkin puree, carrot puree, or sweet potato puree in place of the butternut squash puree.
Even adults love this simple healthy quesadilla when paired with a Fresh Cilantro Lime Cream.
Mix together sour cream (or Plain Greek Yogurt) with lime zest, lime juice, salt and fresh cilantro.
A healthy cheese quesadilla filled with cheese and butternut squash.
Spread 4 tortillas with 1/4 cup squash puree each. Top with one ounce of cheese on each. Top with remaining quesadilla and then heat in skillet, oven, or microwave until cheese is melted.
Mix together the yogurt (or sour cream), cilantro, lime zest and juice and salt. Serve with quesadillas.
Instead of Butternut Squash Puree, you can use sweet potato puree, carrot puree, or even 100% canned pumpkin.
Ooh yes – hello lunch! I've got a huge butternut squash sitting on my counter begging to be used and this sounds perfect!
Yes, this would be a great way to use it up!
This is such a great idea for a quick lunch!
Great idea to bake the butternut squash whole and then cut it! My kids love quesadillas. This would be a hit in my house!!
Yes, I bet they would be!
Kristen, my daughter and your daughter would have so much to share about their EPIC love of cheese and crackers!
I think your daughter and my daughter would probably make pretty epic friends as well!
What a fun idea for kiddos (or husbands)!! Love this!
Yes, fun for husbands and kids for sure Melissa!
Looooove the cheesy butternut pairing in these quesadillas! Totally tasty!
It is surprising how much the butternut squash pairs well with the squash. So glad you enjoyed Jenn!
I love making recipes like this to help the kids get extra veggies! Such a great idea and a delicious recipe!
Thanks so much Meagan! It is a great way to enjoy veggies for sure!