Quantification of microglial late reaction to stereotactic irradiation of the rat brain using computer-aided image analysis

The role of microglial cells in the late delayed reaction following radiotherapy of brain tumors has not been elucidated. To investigate the late delayed response of microglial cells to radiation, we stereotactically irradiated spherical treatment volumes in the right frontal lobe of rat brains. Doses of 20, 30, 40, and 50 Gy were used in combination with two different collimators. The response of microglial cells at 10 and 19 months after irradiation was determined with Anti-CD 11 b/c (Ox 42) as an immunohistochemical marker. For evaluation of immunostaining, we developed a method using computer-aided image analysis in which the ratio of the area of stained cells to that of nonstained brain tissue is calculated. In addition, quantification of Ox-42+ cells per microscopic field was performed. Animals treated with 30 Gy or more had significantly increased total areas of staining at both time points studied. In contrast, the number of stained cells at 10 months increased significantly only in animals treated with 30 or 40 Gy. Likewise, at 19 months, this number increased significantly only in animals treated with 40 Gy or more. These results indicate that computer-aided determination of the area of stained cells is more sensitive than the counting of stained cells. We have demonstrated that microglial cells respond to stereotactic irradiation in a dose-dependent fashion. The image analysis we employed for this purpose is a systematic method to evaluate immunohistochemical staining.     

The berkeleyamides, amides from the acid lake fungus Penicillum rubrum

We previously reported several novel bioactive hybrid polyketide-terpenoid metabolites from a deep water Penicillium rubrum isolated from Berkeley Pit Lake, Butte, Montana. In this paper we report the structures of four new amides, berkeleyamides A-D (1, 4, 5, 7), isolated from extracts of this fungus. The structures of these compounds were deduced by analysis of NMR data, chemical derivatization, and comparison of their spectroscopic data to those of known compounds.     

Shoulder magnetic resonance imaging findings in manual wheelchair users with spinal cord injury

Objective: To investigate the prevalence of rotator cuff and long head of the biceps pathologies in manual wheelchair (MWC) users with spinal cord injury (SCI).Design: Cross-sectional study.Setting: Outpatient clinic at a tertiary medical center.Participants: Forty-four adult MWC users with SCI (36 men and 8 women) with an average age (SD) of 42 (13) years. SCI levels ranged from C6 to L1; complete and incomplete SCI.Outcome Measures: Participants’ demographic and anthropometric information, presence of shoulder pain, Wheelchair User’s Pain Index (WUSPI) scores, and magnetic resonance imaging findings of shoulder pathologies including tendinopathy, tendon tears, and muscle atrophy.Results: Fifty-nine percent of the participants reported some shoulder pain. The prevalence of any tendinopathy across the rotator cuff and the long head of biceps tendon was 98%. The prevalence of tendinopathy in the supraspinatus was 86%, infraspinatus was 91%, subscapularis was 75%, and biceps was 57%. The majority of tendinopathies had mild or moderate severity. The prevalence of any tears was 68%. The prevalence of tendon tears in the supraspinatus was 48%, infraspinatus was 36%, subscapularis was 43%, and biceps was 12%. The majority of the tears were partial-thickness tears. Participants without tendon tears were significantly younger (P < 0.001) and had been wheelchair user for a significantly shorter time (P = 0.005) than those with tendon tears.Conclusion: Mild and moderate shoulder tendinopathy and partial-thickness tendon tears were highly prevalent in MWC users with SCI. Additionally, the findings of this study suggest that strategies for monitoring shoulder pathologies in this population should not be overly reliant on patient-reported pain, but perhaps more concerned with years of wheelchair use and age.     

Induction of Apoptosis in HT-29 Cells by Extracts from Isothiocyanates-rich Varieties of Brassica Oleracea

Among the vegetables with anti-carcinogenic properties, members of the genus Brassica are the most effective at reducing the risk of cancer. This property may be explained by their principle bioactive compounds, isothiocyanates (ITCs). The aim of this study was to measure the amounts of ITCs in extracts from vegetables of the Brasssica genus and assay them for potency of induction of apoptosis in a colorectal cancer cell line (HT-29). ITCs were determined by the cyclocondensation assay with 1,2-benzenedithiol and induction of apoptosis by assessment of cell viability, caspase-3 activity and DNA fragmentation. Purple cabbage extract showed the highest ITC concentration per gram, fresh weight, followed by black cabbage and Romanesco cauliflower. At ITC concentrations of 7.08 μ g/mL these extracts decreased cell viability and induced caspase-3 and DNA fragmentation at 48h. Brussels sprouts showed the strongest effects on cell viability and caspase-3 activity. Varieties of Brassica Oleracea are rich sources of ITCs that potently inhibit the growth of colon cancer cells by inducting apoptosis. All the extracts showed anticancer activity at ITC concentrations of between 3.54 to 7.08 μ g/mL, which are achievable in vivo. Our results showed that ITC concentration and the chemopreventive responses of plant extracts vary among the varieties of Brassica Oleracea studied and among their cultivars.     

State Firearm Legislation and Nonfatal Firearm Injuries

Objectives. We investigated whether stricter state-level firearm legislation was associated with lower hospital discharge rates for nonfatal firearm injuries.Methods. We estimated discharge rates for hospitalized and emergency department–treated nonfatal firearm injuries in 18 states in 2010 and used negative binomial regression to determine whether strength of state firearm legislation was independently associated with total nonfatal firearm injury discharge rates.Results. We identified 26 744 discharges for nonfatal firearm injuries. The overall age-adjusted discharge rate was 19.0 per 100 000 person-years (state range = 3.3–36.6), including 7.9 and 11.1 discharges per 100 000 for hospitalized and emergency department–treated injuries, respectively. In models adjusting for differences in state sociodemographic characteristics and economic conditions, states in the strictest tertile of legislative strength had lower discharge rates for total (incidence rate ratio [IRR] = 0.60; 95% confidence interval [CI] = 0.44, 0.82), assault-related (IRR = 0.58; 95% CI = 0.34, 0.99), self-inflicted (IRR = 0.18; 95% CI = 0.14, 0.24), and unintentional (IRR = 0.53; 95% CI = 0.34, 0.84) nonfatal firearm injuries.Conclusions. There is significant variation in state-level hospital discharge rates for nonfatal firearm injuries, and stricter state firearm legislation is associated with lower discharge rates for such injuries.Each year from 2005 to 2010, an average of 103 000 Americans were injured or killed by a firearm—approximately 282 individuals per day.1 Most public attention and research has focused on fatal firearm injuries because they are a leading cause of injury death and account for more than 30 000 deaths annually.1 Firearm injuries were the third leading cause of injury-related deaths in 2010 after poisoning and motor vehicle accidents and were the second most frequent cause of traumatic death related to a consumer product.1,2 However, a majority of firearm-related injuries in the United States are nonfatal.1,3,4 In 2010, nearly 5 individuals suffered nonfatal firearm injuries for every 2 who died as a result of firearm violence.1 The age-adjusted fatal firearm injury rate in that year was 10.1 per 100 000 person-years, less than half of the age-adjusted nonfatal injury rate (24.0 per 100 000 person-years).1 This high prevalence of nonfatal firearm injuries in the United States is associated with significant physical and psychological morbidity among injury survivors.5–7 It is also a substantial economic burden for victims, taxpayers, and the United States.8–12Numerous state and federal laws have been implemented in attempts to reduce firearm-related violence in the United States.13 Household firearm ownership rates have been shown to be associated with states’ rates of firearm-related suicides and homicides; thus, legislation might reduce firearm injuries by limiting overall firearm ownership.14,15 A strong association has also been demonstrated between safer firearm storage practices and a lower risk of suicide and unintentional firearm deaths.16–19 Hence, legislation aimed at increasing safe firearm storage may decrease firearm-related injuries, particularly in homes with children and adolescents. In addition, laws that promote background checks before firearm purchase and those that limit private firearm transactions and transfers may help limit firearm access by those most likely to harm themselves or others.Overall, the effectiveness of these laws individually or as a whole remains unclear. Two studies evaluated the relationship between state firearm legislation and firearm injuries using measures of state firearm legislation on the basis of annual scorecards created by the Brady Center to Prevent Gun Violence.20,21 These studies found lower rates of total firearm deaths, including homicides and suicides,20 as well as lower rates of firearm injuries in children,21 in states with more restrictive firearm legislation. Previous studies have also shown that laws related to background checks and limitations on handgun possession and transfer are associated with lower rates of firearm deaths, including suicides and homicides.22–25 A study of the 1994 Brady Handgun Violence Prevention Act (Pub L. No. 103-159, 107 Stat. 1536, USC 921–922, HR 1025, 103rd Congress), which established a mandatory waiting period and background check requirement for handgun sales through licensed firearm dealers, found that the law led to a decline in the suicide rate for those aged 55 years and older, although these findings may have been driven by the implementation of the waiting period rather than the background check itself.26Laws focused on preventing children’s access to firearms are associated with lower rates of both unintentional deaths and suicides.27,28 A cross-sectional, time series analysis of pooled data from 1979 to 2000 found that unintentional firearm deaths among children were declining nationally and that most states that enacted child access laws experienced greater declines in those injuries than did states that had not.29 Notably, state-level comparisons of child access laws can be driven largely by the few states with the strictest child access legislation (e.g., felony conviction for violations).29,30Additionally, several studies suggest that laws aimed at easing access to and use of firearms may be associated with higher rates of firearm injuries, including homicides.31–33 One study found that the 2007 repeal of Missouri’s permit to purchase law requiring firearm purchasers to obtain a license verifying that they passed a background check led to an increase in firearm-related homicides.33 Conversely, other studies have observed no association between stricter firearm laws and firearm violence,26,34,35 and a recent systematic review of various federal and state firearm laws found insufficient evidence to determine their effectiveness in reducing firearm-related violence and injuries.36Several studies have examined the relationship between firearm legislation and fatal firearm injuries, although little is known about the relationship between firearm legislation and nonfatal firearm outcomes.37 This relationship may differ from that observed with fatal injuries because of the different circumstances under which nonfatal firearm injuries occur, including differences in the age of the injured,1,3 the type of firearms involved,38 and injury intent.3 For instance, unintentional shootings are more likely to prove nonfatal than are intentional shootings, and a vast majority of self-inflicted injuries (i.e., suicide attempts) result in death.3,39 Because of the higher prevalence of nonfatal firearm injuries, studies of nonfatal injuries may also have greater statistical power to determine associations between legislation and firearm outcomes that might not be observed in studies of fatal injuries.We have described state variation in discharge rates for nonfatal firearm injuries in 2010 and determined whether stricter state-level firearm legislation was associated with lower discharge rates for nonfatal firearm injuries.     

Microbiota That Affect Risk for Shigellosis in Children in Low-Income Countries

Pathogens in the gastrointestinal tract exist within a vast population of microbes. We examined associations between pathogens and composition of gut microbiota as they relate to Shigella spp./enteroinvasive Escherichia coli infection. We analyzed 3,035 stool specimens (1,735 nondiarrheal and 1,300 moderate-to-severe diarrheal) from the Global Enteric Multicenter Study for 9 enteropathogens. Diarrheal specimens had a higher number of enteropathogens (diarrheal mean 1.4, nondiarrheal mean 0.95; p<0.0001). Rotavirus showed a negative association with Shigella spp. in cases of diarrhea (odds ratio 0.31, 95% CI 0.17–0.55) and had a large combined effect on moderate-to-severe diarrhea (odds ratio 29, 95% CI 3.8–220). In 4 Lactobacillus taxa identified by 16S rRNA gene sequencing, the association between pathogen and disease was decreased, which is consistent with the possibility that Lactobacillus spp. are protective against Shigella spp.–induced diarrhea. Bacterial diversity of gut microbiota was associated with diarrhea status, not high levels of the Shigella spp. ipaH gene.