The dramatic success of insecticide treated nets (ITNs) and long-lasting insecticidal nets (LLINs) in African countries has been countered by the rapid development of pyrethroid resistance in vector mosquitoes over the past decade. One advantage of the use of pyrethroids in ITNs is their excito-repellency. Use of the excito-repellency of pyrethroids might be biorational, since such repellency will not induce or delay the development of any physiological resistance. However, little is known about the relationship between the mode of insecticide resistance and excito-repellency in pyrethroid-resistant mosquitoes.An. gambiae s.s., An. arabiensis, and An. funestus s.s., and laboratory colonies of An. gambiae s.s. and An. arabiensis. The contact repellency to pyrethroids or permethrin-impregnated LLINs (Olyset® Nets) was evaluated with a simple choice test modified by WHO test tubes and with the test modified by the WHO cone bioassay test.Differences in the reactions of 3 major malaria vectors in western Kenya to pyrethroids were compared in laboratory tests. Adult susceptibility tests were performed using World Health Organization (WHO) test tube kits for F1 progenies of field-collected An. gambiae s.s., An. arabiensis, and An. funestus s.s. showed high resistance to both permethrin and deltamethrin. The allelic frequency of the point mutation in the voltage-gated sodium channel (L1014S) in An. gambiae s.s. was 99.3–100%, while no point mutations were detected in the other 2 species. The frequency of takeoffs from the pyrethroid-treated surface and the flying times without contacting the surface increased significantly in pyrethroid-susceptible An. gambiae s.s. and An. arabiensis colonies and wild An. arabiensis and An. funestus s.s. colonies, while there was no significant increase in the frequency of takeoffs or flying time in the An. gambiae s.s. wild colony.Field-collected An. gambiae s.s. than in An. arabiensis and An. funestus s.s. It might be that resistant mosquitoes governed by knockdown resistance (kdr) loose repellency to pyrethroids, whereas those lacking kdr maintain high repellency irrespective of their possessing metabolic resistance factors to pyrethroids. Further genetic evaluation is required for the demonstration of the above hypothesis.A different repellent reaction was observed in the field-collected  Pyrethroids are the predominant insecticides and are used in various formulations for mosquito control. Globally, pyrethroids comprise 40% of the insecticides used annually for indoor residual spraying against malaria vectors. Since tkdr). The second is metabolic resistance that relates to the elevated activity of 1 or more detoxification enzymes . The distribution patterns of the types of point mutations in voltage-gated sodium channels in African malaria vectors are unique[Anopheles gambiae Giles (An. gambiae sensu stricto [s.s.]). L1014F mutations are widespread in western African countries, L1014S mutations are distributed in eastern Africa, and L1014F/L1014S hybrids in An. gambiae s.s. are reported in the central region. In contrast, such kdr mutations do not seem to be common in An. arabiensis Patton. One plausible explanation for such a species-dependent difference in host feeding activities under LLIN use might be the difference in the repellency to pyrethroids observed in the present report. The historical population decline of An. gambiae s.s. reported by Bayoh et al.[An. gambiae s.s. population. Anopheles gambiae s.s. might have not changed its characteristics as a “midnight feeder” because of the lack of repellency despite the existence of LLINs, and it still has to rely on the limited human blood sources, most of which are protected by LLINs, resulting in the decline in their population size.Chandre et al. and Corbm (>90%). Recentlh et al. might alAn. gambiae s.s. than in An. arabiensis and An. funestus s.s. It might be that resistant mosquitoes governed by knockdown resistance (kdr) loose repellency to pyrethroids, whereas those lacking kdr maintain high repellency irrespective of their possessing metabolic resistance factors to pyrethroids. The comparative studies of the repellency of mosquitoes have mainly studied kdr, and those studying enhanced metabolic factors have been unexpectedly few. Further genetic evaluation should be required to demonstrate the above hypothesis.A different repellent reaction was observed in the field-collected The authors declare that they have no competing interests.HK and KO designed the study, carried out the experiments, and drafted the manuscript. GOD and GS arranged the field studies, obtained informed consents from the participants for the study, and organized the staff for the experiments. SMN, CM, and NM critically revised the protocol for the study. All authors read and approved the final version of the manuscript.


In spite of the numerous prostate cancer (CaP) intervention programmes that have been implemented to address the disparities experienced by black men, CaP prevention, risk reduction, and early detection behaviours remain low among black men. The lack of formal theoretical frameworks to guide the development and implementation of interventions has been recognised as one of the primary reasons for the failure of health interventions. Members of the Florida Prostate Cancer Health Disparity (CaPHD) group employed the Personal Model of Prostate Cancer Disparity (PIPCaD) model and the Health Communication Process Model to plan, implement, and evaluate an intervention programme, the ‘Working through Outreach to Reduce Disparity (W.O.R.D. on Prostate Cancer)’ video for black men. The location for the video was in a barbershop, a popular setting for the targeted group. The video starred CaP survivors, CaP advocates, a radio personality, and barbers. In addition, remarks were provided by a CaP scientist, a urologist, a CaP advocate, a former legislator, and a minister.The W.O.R.D. video was developed to assist black men in meeting the Healthy People 2020 goal for the United States of America. The efficacy of the W.O.R.D. video was successfully established among 143 black men in Florida. Exposure to the video was found to statistically increase CaP knowledge and intention to participate in CaP screening. Furthermore, exposure to the video statistically decreased participants’ perception of the number of factors contributing to decision, uncertainty about CaP screening. Participants were highly satisfied with the video content and rated the quality of the video to be very good. Participants also rated the video as credible, informative, useful, relevant, understandable, not too time consuming, clear, and interesting. The American Cancer Society (ACS) estimated that there will be about 233,000 new cases of prostate cancer (CaP) in the United States (US) in 2014, and about 29,480 men will die from the disease [The personal factors of black men that have been linked to CaP disparity include low CaP knowledge –7, low Chttps://www.youtube.com/watch?v=74NPk6u_wcw), a prostate cancer education video, designed to close the CaP disparities gap between black men and white men.Based on over 10 years of research, the Florida Prostate Cancer Health Disparity Research (CaPHDR) group developed a Personal Integrative Model of Prostate Cancer Disparity (PIPCaD) for black men –6, 19–23In line with our long-term goal of improving prostate health in minority, rural, and under-served communities, the W.O.R.D video focuses on explaining the risk factors for CaP, how to reduce the risk for CaP, and informed decision making about CaP screening. The W.O.R.D video addressed the limitations of previous CaP intervention programmes for black men using two innovative strategies. The first strategy was the use of a theoretical framework, the PIPCaD model, to plan and develop a video. The PIPCaD model provided information on the behavioural factors to target the CaP education intervention, including: attitude, perceived severity, perceived susceptibility, cues to action, knowledge, and perceived behavioural control. The CaP screening facilitators and deterrents targeted are summarised in The second strategy is the use of the Health Communication Process Model  to ensurThe primary goal of this study was to develop a video intervention that will improve the prostate health behaviour of black men. To accomplish this goal, the study objectives were to:Develop a culturally relevant CaP education intervention, the W.O.R.D video, for black men based on the PIPCaD model.Evaluate participants’ general assessment of the W.O.R.D video, including satisfaction with and perceived quality of the video.Establish the efficacy of the W.O.R.D video in improving users’ CaP knowledge and CaP screening intention as well as reducing decisional conflict about CaP screening.The intervention programme was based on the PIPCaD model, which provided the understanding on the needs and perceptions of black men. The strategy statement for the intervention programme see  was deveThe primary intervention channel for the W.O.R.D programme is the video. Video has been found to be one of the most effective channels for educating black men , 26. It The intervention sources were primarily characters that black men can relate to and included: (1) Malik, the barber, a young black man; (2) Mr King, a 62-year-old customer, on his way to get his first prostate check-up; (3) Andre, an older customer, who has been screened regularly for years; (4) Quincy, a mid-40s customer with a comedic personality; (5) Chuck, a CaP survivor; and (6) set extras who were barbers and young customers.The modifiable variables targeted for the intervention message were: (1) perceived behavioural control; (2) perceived susceptibility to CaP; (3) perceived severity of CaP; and (4) cues to action. A video and multimedia company was hired to produce and direct the video, including pre-production meetings and correspondence with the research team, which included CaP survivors; scriptwriting; video production planning; video shoot of all scenes; and post-production, including editing into final video sequences for website and Digital Video Disc (DVD). The storyline for the script was that Quincy (a black man in his mid-40s) becomes aware of the importance of CaP prevention and having a consultation with a physician for informed decision on screening, and finally decides to discuss it with his doctor after a lively conversation with his friends at a barbershop. A knowledgeable friend and a CaP survivor share key information with Quincy, the barbers, and other customers. While the storyline was anchored on humor, it highlighted specific messages on risk reduction and early detection of CaP. At the end of the video, the CaP message was reinforced by scrolling on the screen and simultaneously announcing information about the CaP risk factors, specific steps that can be taken to reduce an individual’s risk for cancer, the pros and cons of CaP screening, and how to make an informed decision about CaP screening.The study was a pre-, post-test research design, exploring the impact of the W.O.R.D video on the following variables: CaP knowledge, CaP screening intention, and decisional conflict. These three variables were selected as the study variables because they have been found to impact CaP risk reduction and screening behaviours –10. TablThe study setting for this project was Alachua and Orange counties in Florida. The inclusion criteria were black men, age 35 and above, regardless of CaP history. Men who were not black men of African ancestry and who were unable to speak English were excluded from participating in the study. Participants were recruited at health forums and community outreach events, including the African American Men’s Summit in Orlando, the Men’s Health Expo in Gainesville, and the University of Florida mobile health clinics in Gainesville. Participants were recruited by undergraduate research assistants, including the University of Florida-Florida A&M University ReTOOL students as part of their CaP research training experience. The research assistants approached black men at these events and asked if they would like more information about the study or have any questions about participating in the study. Only the men who indicated an interest in participating in the study were included after verbal consent.Institutional Review Board (IRB) approval for the project was received on 29 March 2012. Subsequently, data were collected prospectively from participants between 1 April 2012 and 16 June 2012. Data collection included the following three steps:Step 1: Participants completed a self-administered pre-test survey measuring CaP knowledge, CaP screening intention, decisional conflict, medical access information, and demographic information.Step 2: Participants viewed the W.O.R.D video. The video was viewed in groups of 5–10 depending on the study site.Step 3: Following the viewing of the W.O.R.D video, participants completed a self-administered post-test survey measuring CaP knowledge, CaP screening intention, decisional conflict, satisfaction with the W.O.R.D video, perceived quality of the W.O.R.D video, and general assessment of the W.O.R.D video.The survey instructions noted that participants may choose not to answer all of the survey questions and they may withdraw from the study at any time. During data collection, participants who asked for assistance in completing the survey, mostly due to difficulty in reading, were assigned a research assistant to assist them in completing the survey. A US$10–15 Wal-Mart gift card was provided to all participants as an incentive for their participation.The survey data were coded and stored in a Microsoft (MS) Excel database. Subsequently, Psychological Clinical Science Accreditation System (PC-SAS) analytical software was employed for data cleaning and analyses. Frequency analysis was first conducted to confirm that the responses were appropriately entered and to correct any errors. The statistical analyses included: internal consistency of the study scales to establish the reliability of the measures; descriptive statistics to summarise socio-demographic, and study variables; and T-test analyses to check the changes in key study variables between pre- and post-intervention exposure.Our primary goal was to develop an education intervention programme that will improve the prostate health behaviour of black men and establish the efficacy of the programme. Based on the Health Communication Process , we accohttp://www.youtube.com/watch?v=74NPk6u_wcw). The final version of the W.O.R.D video was produced on 28 February 2012. Two versions of the video were produced. The full version is about 25 minutes long and the short version is about 14 minutes long. The short version is hosted on YouTube  it iStudent’s T-test analysis was conducted to compare the differences between the means of the pre-test scores and post-test scores on CaP knowledge, intention to screen for CaP, decision uncertainty and uncertain factors after the intervention exposure. A summary of the T-test results is provided in Black men were the only racial group that did not meet the CaP Healthy People 2010 goal for the US to reduce the CaP death rate to 28.8 deaths per 100,000 males by 2010 . BetweenThe W.O.R.D. video was developed to assist black men in meeting the Healthy People 2020 goal for the USA. The efficacy of the W.O.R.D. video was successfully established among 143 black men in Florida. Exposure to the W.O.R.D. video was found to statistically increase CaP knowledge and intention to participate in CaP screening. Furthermore, exposure to the W.O.R.D. video statistically decreased participants’ perception of the number of factors contributing to decision uncertainty about CaP screening. Participants were highly satisfied with the video content and rated the quality of the video to be very good. Participants also rated the video as credible, informative, useful, relevant, understandable, not too time consuming, clear, and interesting.It is important to note that the benefits of CaP screening for the general population remain controversial. However, given the disproportionate burden of CaP in black men and the lack of clinical trials to assess the effectiveness of screening in black populations, the only strategy we currently have to combat CaP disparity for black men is the early detection of CaP. The W.O.R.D. video promotes informed decision making by black men, listing the pros and cons of CaP screening.This project only focused on the short-term impact of the W.O.R.D. video, focusing on its efficacy and not the effectiveness in real life settings. The next step would be to assess the effectiveness and uptake of the intervention. While there are several advantages to providing CaP education at barbershops, as pointed out above, it is important to note that this cannot serve as a surrogate to talking with a healthcare provider. The W.O.R.D. video intervention is likely to be more effective in barbershops if the barbers are trained to be prostate health advisers. In the absence of trained barber health advisers, the barbers will need to navigate their clients to appropriate health care providers for prostate health education. Another limitation is the representation of participants. Since participants were not recruited randomly and constitute a convenient sample, they are not representative of all black men. As mentioned above, the participants for the study were more knowledgeable about CaP and had a higher CaP screening intention because of their exposure to CaP information at health forums. In spite of these limitations, the W.O.R.D. video has significant advantages. It was tailored to the specific needs and characteristics of black men because it was developed from information generated from black men on their CaP personal factors. The W.O.R.D. video message is thus credible, relevant, and culturally-responsive to black men. The W.O.R.D. video is also highly accessible nationwide through YouTube, fostering widespread dissemination, at minimal cost. In addition, the W.O.R.D. video DVD and accompany brochures are available at no charge to black men.There is no conflict of interest reported by authors.


Rickettsia spp. belonging to the spotted fever group (SFG) cause infections in humans, domestic animals and wildlife. At least five SFG rickettsial species have been reported in China, but the occurrence of Rickettsia aeschlimannii and R. massiliae in ticks has not been characterized to date.Rickettsia spp. in the ticks. The ticks collected were identified as Rhipicephalus turanicus. Three different lineages of Rh. turanicus from Yining County were discovered on phylogenetic analysis of 16S rDNA and cox1. Twenty-one of the 114 samples (18.42%) were positive for rickettsial agents. Phylogenetic analysis based on six genetic sequences showed that three rickettsial species were present, namely: R. aeschlimannii , R. massiliae  and R. sibirica variant , which is clustered in the clade of R. sibirica subsp. sibirica.A total of 114 adult ticks were collected from sheep in Yining County, Xinjiang Uygur Autonomous Region, in northwest China. The ticks were identified from morphological and molecular characteristics. All samples were examined by polymerase chain reaction (PCR), and six genetic markers were used to determine the R. aeschlimannii and R. massiliae in China. R. massiliae, R. aeschlimannii and R. sibirica variant co-circulate in the region of the China-Kazakhstan border, in northwest China. Rickettsial agents in ticks of the genus Rhipicephalus from migrant birds, transported livestock, wildlife and human beings should be investigated further in the region of the China–Central Asian border.This is the first description of The online version of this article (doi:10.1186/s13071-015-1242-2) contains supplementary material, which is available to authorized users. Rickettsia spp. belonging to the spotted fever group (SFG) cause infections in animals and humans worldwide  were amplified using previously described primers .Twenty-one of the 114 samples 18.42%) were positive by PCR for products of six rickettsial genetic markers. Out of the 21 positive samples, four were confirmed as % were po% 722bp/71bp and 9R. massiliae, R. rhipicephali and R. aeschlimannii are grouped phylogenetically into a clade in the family Rickettsiaceae [R. massiliae was first isolated in 1990 from a Rh. turanicus tick in an area near Marseille, France [Rhipicephalus ticks in regions of Europe, North and Central Africa, and the United States [R. aeschlimannii was first described from Hyalomma marginatum in Morocco in 1997 [R. aeschlimannii has been demonstrated in Hyalomma ticks from Europe , Asia  and Africa  [Haemaphysalis ticks in Spain and Kazakhstan [Ixodes ricinus, H. punctata, Rh. bursa, and Rh. sanguineus isolated from human Spanish patients were shown to contain DNA from R. aeschlimannii [R. aeschlimannii from Rh. turanicus infecting a man in Greece in 2013 [R. aeschlimannii and R. massiliae are present in Rh. turanicus from sheep in the region of the China-Kazakhstan border, in the northwest of China.tsiaceae . R. mass, France . Since td States . Further in 1997 . The pre, Egypt) –16 and fzakhstan . Furtherlimannii , and the in 2013 . In thisR. sibirica is known to contain two subspecies [R. sibirica subsp. sibirica and R. sibirica subsp. mongolotimonae. The former was first isolated in Russia but it has subsequently been found in northern China [R. sibirica subsp. mongolotimonae was first isolated in Inner Mongolia and then found in Europe and Africa [R. sibirica variant found in the region of the China–Kazakhstan border appeared divergent in the ompA, ompB and sca1, used to differentiate Rickettsia species, although it was closest to R. sibirica subsp. sibirica, on the basis of the gltA gene and the phylogenetic tree of the 17-kDa-ompA-gltA-rrs-sca1-ompB concatenated sequence. Further genomic analysis should be carried out to confirm the classification of the R. sibirica variant found in this study.To date, bspecies , R. sibirn China . In contd Africa , 21. HerRh. turanicus tick is widely distributed throughout the Mediterranean subregion, Africa, and Asia, including China, especially in XUAR [Rickettsia spp. [R. massiliae, R. aeschlimannii and R. sibirica variant were found in the same area, Yining County, which suggests that several SFG Rickettsia spp. co-circulate in Rh. turanicus as a potential vector near the China-Kazakhstan border.The  in XUAR , and it sia spp. . Here, Ret al detected R. aeschlimannii in the Alma-Ata region, east of Kazakhstan [Rh. turanicus in the region of the China-Kazakhstan border showed genetic divergence in the loci of 16S rDNA and cox1, which indicates that these ticks collected from sheep may come from different lineages. At present, it is unknown whether these ticks are imported from the Chinese hinterland or abroad through migrant birds, or with internationally transported livestock. This topic needs to be further investigated.In 2004, Shpynov zakhstan . Here weR. aeschlimannii and R. massiliae in China. The findings of the study suggest that R. massiliae, R. aeschlimannii and R. sibirica variant co-circulate in Rh. turanicus in the region of the China–Kazakhstan border, in northwest China. The origin of the Rhipicephalus genus  and the epidemiology of tick-borne pathogens should be further investigated in the region of the China–Central Asian border.This is the first report of the molecular analysis of


Objectives. This study aimed to assess the effect of hydroalcoholic extract of Punica granatum Linn. (P. granatum) petal on Streptococcus sanguinis, Streptococcus mutans, Streptococcus salivarius, Streptococcus sobrinus, and Enterococcus faecalis. Materials and Methods. In this in vitro study, P. granatum extract was prepared using powdered petals and water-ethanol solvent. Antibacterial effect of the extract, chlorhexidine (CHX), and ampicillin was evaluated on brain heart infusion agar (BHIA) using the cup-plate method. By assessing the diameter of the growth inhibition zone, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined for the above-mentioned bacteria. Results. Hydroalcoholic extract of P. granatum petal had inhibitory effects on the proliferation of all five bacterial strains with maximum effect on S. mutans with MIC and MBC of 3.9 mg/mL. The largest growth inhibition zone diameter belonged to S. sanguinis and the smallest to E. faecalis. Ampicillin and CHX had the greatest inhibitory effect on S. sanguinis. Conclusions. Hydroalcoholic extract of P. granatum had a significant antibacterial effect on common oral bacterial pathogens with maximum effect on S. mutans, which is the main microorganism responsible for dental plaque and caries. Dental caries, periodontal disease, and opportunistic infections are among the common diseases of the oral cavity caused by the accumulation of pathogenic microorganisms, inappropriate nutritional habits, and improper oral hygiene –4. Oral Herbal medicine has a long history. Many individuals residing in developing countries strongly believe in the efficacy of herbal medications for primary care , 13. Con Punica granatum Linn. is a fruit-bearing deciduous shrub or small Asian tree from the family of Punicaceae growing between 5 to 8 meters tall. It is native to Iran and northern India and has long been cultivated in the Asian Mediterranean region, Europe, and Africa. Punica granatum is rich in bioactive compounds, which are used for treatment of cancer, cardiovascular diseases, diabetes mellitus, dental diseases, bacterial infection, antibiotic resistance, skin conditions due to UV radiation, diarrhea, bloody diarrhea, and hemorrhoids. Moreover, P. granatum is used as a mouth rinse for treatment of some types of sore throat. Parts of the plant used for medicinal purposes include its flowers, trunk skin, fruits, roots, and seeds [The pomegranate with the botanical name ofnd seeds –21. Punica granatum flowers are odorless but colorful red or reddish, 3.5 to 7 cm in length, and campanulate or cylindrical. Flowers are two types: fertilized and unfertilized. The unfertilized flower is with smaller, barren, and short-styled, short-stamened petals, in which the stigma is far below the anthers. The unfertilized flowers are commonly known as “Golnar” in Iranian traditional and complementary medicine [Themedicine .Trunk skin and roots are used for parasitic infections, dried flowers are beneficial for treatment of bronchitis, diarrhea, and bloody diarrhea, and the brewed pomegranate is used for treatment of inflammation of the throat and oral cavity. Pomegranate has long been recommended as a hemostatic agent and for treatment of diabetes mellitus in ancient Greek medicine –21. P. granatum have been recently noticed [ P. granatum have shown strong antimicrobial properties against Gram-positive and Gram-negative nonoral microorganisms [The antimicrobial properties of noticed , 23–30. rganisms , 24, 31.rganisms , 31–33. P. granatum flower  on oral microbial pathogens, which showed its greatest antimicrobial effect on S. sanguinis [ P. granatum was very effective against biofilm forming microorganisms in the mouth of patients [However, the mentioned studies are very limited and mostly lack an appropriate microbiological method. Their methodology is often not clear or not well described. Search of the literature only yielded one previous study on the effect of water extract ofanguinis , althougpatients . P. granatum petal on S. mutans, S. sanguinis, S. salivarius, S. sobrinus, and E. faecalis in vitro.Considering the high prevalence of oral and dental diseases due to oral pathogens and the recent public interest in medicinal plants, this study aimed to assess the effect of hydroalcoholic extract ofP. granatum flowers were obtained from the last harvest in Darab city and its purity was confirmed in a pharmacognosy laboratory. P. granatum extract was prepared by maceration method. Powdered petals were precisely measured by a digital scale and poured into an Erlenmeyer flask. Hydroalcoholic solvent (50% water and 50% ethanol) was also added. The Erlenmeyer flasks were capped with aluminum foil and stored in the dark for 10 days. Next, the flasks were placed on a shaker (GFL 3017) operating at 90 rpm for 24 hours. The solutions were then paper filtered. The filtered solution was poured into a sterile glass container and capped by aluminum foil. A few holes were perforated in the foil and the glass container was placed in Bain-Marie at 90°C to dry. The dried extract was precisely weighed, labeled, and refrigerated [igerated . S. mutans , S. sanguinis , S. salivarius , S. sobrinus (ATCC 27607 and PTCC 1601), and E. faecalis  were obtained in lyophilized form from the Persian Type Culture Collection center. Bacteria were activated by inoculation in the brain heart infusion agar  culture medium followed by 24 hours of incubation at 37°C. For preparation of microbial suspension, a 24-hour culture was used. The concentration of microorganisms in the microbial suspension was adjusted to 0.5 McFarland standard using a spectrophotometer at a wavelength of 625 nm  [Standard strains ofμL of each microbial suspension at 0.5 McFarland standard concentration was cultured in BHIA (swabbed on the plate). Then, wells measuring 8 mm in diameter were created on the agar surface. Different concentrations of the extracts were prepared by serial dilution  using sterile distilled water solvent; 100 μL of each concentration of extract was poured into each well. The plates were incubated at 37°C  for 24 hours. The diameter of the growth inhibition zone was measured in millimeters. This process was repeated in triplicate and the mean diameter of the growth inhibition zone was calculated for different concentrations of the extract [Primary assessment of the antimicrobial effect of extracts was done using the cup-plate technique. 500  extract .6 CFU/mL. Microbial suspension was then diluted by one-half using the culture medium and 1 mL of it was added to the tubes containing serially diluted extract. The negative control tube only contained the culture medium and extract with no microbial suspension. The positive control tube contained culture medium and microbial suspension with no extract. After 24 hours of incubation at 37°C, growth and proliferation of microorganisms were evaluated and the MIC value of the extract for each bacterial strain was determined. This test was repeated in triplicate for each microorganism [The MIC is defined as the lowest concentration of an antimicrobial agent that inhibits the growth of a microorganism (0.5 McFarland standard in this study). It is the minimum concentration of the extract that completely prevents visible growth and proliferation of bacteria compared to the negative control group. To determine MIC, macrodilution method according to the standard technique described by the clinical and laboratory standards institute (2012) was used. Different concentrations of the extract were prepared by serial dilution  in BHI broth medium. Using this medium, the 0.5 McFarland standard suspension was diluted 1 to 150 to obtain a bacterial count of 10organism .μL of the suspension in the tube containing MIC of the extract and tubes showing no bacterial growth were cultured on plates containing BHIA. After 24 hours of incubation at 37°C, the plates were evaluated for growth of microorganisms. The concentration with no bacterial growth was determined as MBC. This test was repeated in triplicate for each microorganism [After determination of MIC, 20 organism .The effect of 0.2% CHX  and ampicillin on the microorganisms was also evaluated using the cup-plate method and the MIC and MBC values of CHX and ampicillin for the microorganisms were determined as well.The tests were repeated in triplicate and the mean and standard deviation (SD) of the growth inhibition zone diameter in cup-plate method as well as the MIC and MBC of the extract, CHX, and ampicillin were determined. P. granatum showed inhibitory effects on the growth and proliferation of all five bacteria using the cup-plate method. The largest and the smallest diameter of growth inhibition zone belonged to S. sanguinis and E. faecalis, respectively. The mean (and SD) diameter of the growth inhibition zone due to the effect of hydroalcoholic extract of P. granatum on different microorganisms is shown in The hydroalcoholic extract of P. granatum were determined using serial dilution method. The highest antibacterial effect of P. granatum extract was on S. mutans with MIC and MBC values of 3.9 mg/mL. The MIC and MBC values of this extract for different microorganisms are presented in Tables The MIC and MBC of hydroalcoholic extract ofThe cup-plate method showed that 0.2% CHX had antimicrobial effect on all five bacterial strains. The mean and SD of the diameter of growth inhibition zone due to the effect of 0.2% CHX on the five bacterial strains are shown in  S. sanguinis. Tables Assessment of the MIC and MBC values revealed that CHX had the highest effect on S. mutans and S. sanguinis due to the effect of ampicillin. The mean and SD of the diameter of growth inhibition zone due to the effect of ampicillin on different microorganisms are shown in The cup-plate method showed that ampicillin had inhibitory effects on the growth and proliferation of all five bacterial strains. The greatest diameter of growth inhibition zone belonged to S. sanguinis. Tables Assessment of the MIC and MBC values revealed that ampicillin had the highest effect on P. granatum petal on S. mutans, S. sanguinis, S. salivarius, S. sobrinus, and E. faecalis in vitro using the cup-plate method. The MIC, MBC, and growth inhibition zone diameter values of the extract for different microorganisms were also calculated and compared to CHX and ampicillin.This study was the first to experimentally assess the antimicrobial effect of hydroalcoholic extract of P. granatum petal had inhibitory effect on the growth and proliferation of all five bacterial strains. Vasconcelos et al. evaluated the MIC of pomegranate fruit gel and compared it with miconazole against adhesion of S. mutans, S. mitis, and C. albicans. They showed that pomegranate fruit gel was more effective than miconazole in preventing the adhesion of streptococci to glass [ P. granatum peel (MEPGP) was effective against L. acidophilus, S. mutans, and S. salivarius. Furthermore, no concentrations of MEPGP inhibited A. viscosus and C. albicans [ P. granatum petal was effective against S. mutans in lower concentration than MEPGP in Abdollahzadeh et al. study. Therefore, it seems that hydroalcoholic extract of petal against S. mutans was more effective than MEPGP. It should be considered that S. mutans is main microorganism of dental plaque and caries [ S. mutans were similar (3.9 mg/mL).Based on the results, the hydroalcoholic extract ofto glass . Dahham to glass . Howeveralbicans . In presd caries . Also, i P. granatum showed the best antibacterial effect of this extract on S. mutans, while the lowest effect was noticed on E. faecalis. The only previous study on the antimicrobial effect of methanolic extract P. granatum flower on S. mutans was conducted by Haghighati et al. [ S. mutans. Also, they showed no significant difference between the efficacy of P. granatum and CHX against S. mutans; these findings show that the method used was not proper and therefore their results are not reliable.The MIC and MBC values ofi et al. . Their r P. granatum flower  on the same microorganisms as in our study was evaluated. The highest antimicrobial effect was seen on S. sanguinis [ S. mutans.In a study by Vahid-Dastjerdi et al., the effect of water extract ofanguinis  comparedP. granatum has well known isolated components such as gamma terpinene, borneol, camphor, carvacrol methyl ether, and methyl palmitate [One important factor affecting the MIC is the difference in the composition of extracts. The composition of extract is influenced by the geographical location of the plant, season of harvesting, age of plant, growth stage, method of drying, and extraction technique. Also, extracts of different parts of the plant have variable level of antimicrobial activity and bacteria have variable sensitivity to different extracts. Also, isolated components of an extract show greater antimicrobial effects than the extract itself . Alcoholalmitate . S. mutans, S. sanguinis, S. salivarius, S. sobrinus, and E. faecalis [ P. granatum with confirmed antimicrobial effects. Thymol is an isomer of carvacrol, showing its antimicrobial activity by making the bacterial membrane permeable [Camphor is one of the main components with antibacterial effect. A previous study isolated the constituents of rosemary and evaluated their effects on oral pathogenic microorganisms. Camphor had inhibitory effects onfaecalis . Carvacrermeable . Chlorheermeable . The MIC P. granatum flowers harvested at a specific time from a specific location. Also, standardized microbial strains were used and microorganisms collected and purified from the oral cavity were not assessed. Thus, the results were limited to standard strains in the laboratory setting. Future studies are required to confirm the results in normal biological environment . Also, future studies must focus on the isolated constituents of different extracts and the antimicrobial agent in their composition. Given that the effects of these constituents on other microorganisms such as L. acidophilus, A. viscosus, and C. albicans are confirmed, they may be used in the form of pure extract, mouth rinse, or other antimicrobial products in clinical trials.One limitation of our study was that we only evaluated the P. granatum had a significant antibacterial effect on common oral bacteria, namely, S. sanguinis, S. mutans, S. salivarius, S. sobrinus, and E. faecalis.Hydroalcoholic extract of P. granatum had maximum antibacterial effect on S. mutans with MIC and MBC of 3.9 mg/mL.Hydroalcoholic extract ofWithin the limitations of this study, the following conclusions were drawn:


Liposcelis bostrychophila, Liposcelis decolor, and Liposcelis paeta. Five adult females of these species were placed in vials containing wheat, either alone or in all possible combinations of two species. The number of adults in the vials was counted after 35, 70, 105, 140, and 175 days. These tests were performed at 25 and 30°C. At 25°C, there were no differences in numbers of L. bostrychophila when this species was reared either alone or with each of the other two species. At 30°C, L. bostrychophila was the dominant species. The presence of L. bostrychophila had a negative effect on the growth of populations of L. decolor and L. paeta. The presence of L. paeta did not affect growth of populations of L. decolor, although the presence of L. decolor occasionally reduced growth of populations of L. paeta. In the second series of tests, L. bostrychophila adult females were placed in vials of wheat either alone or with adult females of Lepinotus reticulatus, at the ratios of (L. bostrychophila: L. reticulatus) 10∶0, 9∶1, 7∶3, 5∶5, 3∶7, 1∶9, and 0∶10. These tests were carried out only at 30°C, and the observation periods were the same as for the first series of tests. Liposcelis bostrychophila was the dominant species in this case as well, regardless of the ratio of the parental females. At the end of the experimental period, L. reticulatus was present only in vials that contained this species alone. Our results showed that L. bostrychophila outcompetes the other stored-product psocid species tested.We evaluated the competition among stored-product psocid species by conducting two series of laboratory experiments. In the first series, three species of Liposcelididae were used:  Tribolium castaneum, increased with the increase of the presence of the lesser grain borer, Rhyzopertha dominica, in the same sampling unit, but the numbers of these two species were negatively correlated with the presence of the rusty grain beetle, Cryptolestes ferrugineus. Athanassiou and Saitanis Ephestia kuehniella, and its parasitoids Harbobracon hebetor and Venturia canescens.The insect community in the stored-grain agroecosystem consists of several categories of species, such as primary and secondary colonizers, fungus feeders, scavengers, predators, and parasitoids There are numerous laboratory studies investigating the competition among different stored-product insect species Liposcelis entomophila in two bins of wheat in 2005, while in the same bins, they found only L. decolor one year later (2006) after fumigating the grain between years.Most of the studies that examine competition among stored-product insect species under laboratory conditions tested beetles, and, to a lesser extent, moths. There are no published data on competition among psocid (Psocoptera) pest species. Psocids, particularly some species of the family Liposcelididae, are major pests that can easily build up high populations, especially in stored grains Despite the fact that coexistence of some major stored-product insect species has been investigated, there are no data available describing competition among species of stored-grain psocid pests. Thus, our study aims to provide the first series of data on the interspecific competition among species of stored-product psocid pests examining the long-term population growth of several psocid species placed alone (one species) or in pairs of species in the same food source.Liposcelis bostrychophila, L. decolor, L. paeta, and Lepinotus reticulatus were used in the tests. L. bostrychophila and L. reticulatus are predominantly parthenogenetic, although a few males of L. bostrychophila were recently found Liposcelis spp. , and also each species alone. For this purpose, 5 adult females of each species were placed in plastic vials  with 5 g of untreated, clean, whole wheat (variety Fuller) adjusted to 13% moisture content. Five adult females were placed in vials where only one species was used. All vials then were placed in incubators set at 30°C and 75% relative humidity. The vials were opened after 35 days, and the adults of the two species were sieved from the wheat using a #30 sieve, identified, and counted. Nymphs were not identified nor counted, given that identification of nymphs of different species of the genus Liposcelis is difficult In these tests, we assessed each possible two-way combination of the three L. bostrychophila and L. reticulatus were used. Ten adult females were placed in each vial at the following combinations of L. bostrychophila: L. reticulatus adult ratios: 10∶0, 9∶1, 7∶3, 5∶5, 3∶7, 1∶9, and 0∶10. The experimental procedure, the conditions, and the observation periods were as described above, with the exception that this experiment was carried out only at 30°C. In these tests, both adults and nymphs were identified and counted, given that all stages of the genera Liposcelis and Lepinotus can be distinguished easily In these tests, For Experiment 1, the data were analyzed, separately for each species, using a three-way ANOVA with temperature, observation period, and psocid species as main effects, with psocid counts as the response variable. Similarly, for Experiment 2, the data were analyzed using a two-way ANOVA, with observation period and species ratio as main effects. All analyses were conducted using the JMP 10 software . Means were separated by the use of the Tukey-Kramer HSD test at α = 0.05 2. All analyses were performed using R version 3.0.2 The evolution of adult counts for the different insect species was modelled as a function of time using a quadratic regression model with temperature and initial counts of insects as covariates. The quadratic term for time was preferable to alternatives given the fact that it allows the possibility of capturing U-shaped patterns and steep rates of decline as time increases L. bostrychophila, all main effects were significant, but only the Period×Temperature interaction was significant  was modeled using a quadratic regression model taking into account the L. bostrychophila: L. paeta: L. decolor initial counts and temperature . The initial number of L. paeta had a significant effect on L. bostrychophila adults counts  and similarly for L. decolor . The effect of temperature was highly significant with lower temperature resulting in higher adult L. bostrychophila counts .The population growth (population evolution) of L. paeta adult counts as a function of time (in days) was modeled as for L. bostrychophila . There was no significant evolution of L. paeta adult counts over time . However, the initial number of L. bostrychophila had a significant effect on L. paeta adult counts , while the same was found for L. decolor . The effect of temperature was highly significant with lower temperature resulting in lower adult L. paeta counts .The population evolution of L. decolor adult counts was similarly modeled . There was no significant evolution of L. decolor adults' counts over time . The initial number of L. bostrychophila had a significant effect on L. decolor adult counts . However, the initial number of L. paeta did not significantly affect L. decolor counts . The effect of temperature was highly significant with lower temperature resulting in higher adult L. decolor counts .Finally, the population of L. bostrychophila adults and nymphs, Period and Species ratio×Period were significant, but Species ratio was not , L. bostrychophila population levels were lower when other species were present. The analysis performed here suggested that despite the fact that this species was always the winner of the competition, its population growth was highly mediated by the presence of other species, which means that competition is likely to slow population increase in a more realistic scenario of a larger spatial scale. This clearly indicates that competition, at least for L. bostrychophila, is highly mediated by temperature, and, at low temperatures, coexistence may occur. In contrast, the adult population of L. decolor was continuously higher when this species was reared alone at 25°C than when this species coexisted with L. bostrychophila. Moreover, there were no differences in L. decolor adult numbers at 30°C for the last 140 days of the observation period. L. paeta adult numbers were reduced at both temperatures when reared with other species. Rees and Walker L. bostrychophila was three times higher than that of L. paeta, but the differences were minimal at lower temperatures. Finally, for both temperature levels tested here, egg-to-adult survivorship was higher for L. bostrychophila than for L. decolor or L. paetaL. bostrychophila at 25°C during the first 35 days of the observation period, as indicated by the high number of adults that were recorded during this period. In contrast, the number of L. paeta adults only increased slightly, and practically remained unaffected for L. decolor.As expected, temperature played an important role in population growth and the concomitant competition. For L. bostrychophila was always the dominant competitor. Again, this species was superior in population growth and life table parameters than L. reticulatus. For example, L. reticulatus egg-to-adult development is slower than that of L. bostrychophilaL. bostrychophila is almost 40 times higher than that of L. reticulatus at 30°C L. bostrychophila over L. reticulatus was manifested regardless of the initial number of L. bostrychophila parental females. Hence, even in the vials that contained only one parental L. bostrychophila female, L. reticulatus became extinct after 140 days. However, the population growth of L. bostrychophila was proportional to the initial number of adults, at least during the first 70 days of the observation period. Interestingly, population growth was inversely related to the initial number of parental adults at the last observation period (175 days). We assume that, as noted above, density-dependent mechanisms regulated single-species population dynamics, given that higher numbers of parental females produced higher populations earlier in the observation period, which resulted in more vigorous intraspecific competition and faster food exhaustion. On the other hand, L. reticulatus population drastically declined very early in the experiment, when L. bostrychophila was present, and this reduction was proportional to the number of parental L. reticulatus females. Eventually, L. reticulatus was present only when it was alone, which suggests that this species has lower population growth rates and does not colonize as well as L. bostrychophila. This species has shorter adult longevity and lower fecundity than L. bostrychophila, which clearly stands in agreement with the data of the current study L. reticulatus was <1% of the total number of psocid individuals recorded.Regarding the second series of experiments, L. bostrychophila was the superior colonizer of the competition, the initial coexistence rate plays an important role in the time that this outcome will occur. In this context, our data documented that coexistence always delays population increase, regardless of the psocid species that coexist. Temperature also plays an important role in this outcome.We used statistical models with linear and quadratic terms for time as predictors in order to capture U-shaped evolutions of counts. Models with more terms or with time added as a predictor with degree higher than 2 were also explored without adding much to the results. We concluded that coexistence plays a significant role in the population growth and probably on the developmental parameters of psocids, regardless of the final outcome of this competition. Hence, despite that, based on our tests, L. bostrychophila was clearly the dominant competitor, but the outcome of this competition was based on the other competitor species, temperature, observation period, and initial population. Overall, our tests show that, under conditions of limited food availability, the simultaneous presence of L. bostrychophila will lead the populations of L. decolor and, especially, L. reticulatus to extinction. Conversely, L. paeta could coexist with L. bostrychophila for a longer period, but in low numbers, which eventually is likely to lead to extinction as well. Further experimentation is needed to assess, apart from population growth, additional factors that are responsible for the superiority of L. bostrychophila over the other species tested here, with emphasis on the possible interactions among individuals, such as possible predation activity among individuals or the existence of semiochemical-based population regulators. The results of the present study will contribute by providing the inferences necessary for the development of long-term prediction plans for psocid colonization and extinction patterns in stored grains and related commodities.To our knowledge, this is the first study in which the inter- and intraspecific competition of stored-product psocid species, expressed as population growth, was assessed, under the basis of limited availability of food or food exhaustion. From the species spectrum tested here,


The Szent-Györgyi Prize for Progress in Cancer Research is a prestigious scientific award established by the National Foundation for Cancer Research (NFCR)—a leading cancer research charitable organization in the United States that is committed to supporting scientific research and public education relating to the prevention, early diagnosis, better treatments, and ultimately, a cure for cancer. Each year, the Szent-Györgyi Prize honors an outstanding researcher, nominated by colleagues or peers, who has contributed outstanding, significant research to the fight against cancer, and whose accomplishments have helped improve treatment options for cancer patients. The Prize also promotes public awareness of the importance of basic cancer research and encourages the sustained investment needed to accelerate the translation of these research discoveries into new cancer treatments. This report highlights the pioneering work led by the 2015 Prize winner, Dr. Frederick Alt. Dr. Alt’s work in the area of cancer genetics over four decades has helped to shape the very roots of modern cancer research. His work continues to profoundly impact the approaches that doctors around the globe use to diagnose and treat cancer. In particular, his seminal discoveries of gene amplification and his pioneering work on molecular mechanisms of DNA damage repair have helped to usher in the era of genetically targeted therapy and personalized medicine. In recognition for his pioneering work in the area of cancer genetics, Dr. Frederick Alt was awarded the 2015 Szent-Györgyi Prize for Progress in Cancer Research. Dr. Alt’s research is foundational to the modern understanding of cancer as a genetic disease, and has led directly to the era of precision medicine. The Prize was presented at a ceremony in Washington, D.C., on April 29, 2015, at The National Press Club.The Szent-Györgyi Prize for Progress in Cancer Research is a prestigious scientific award established by the National Foundation for Cancer Research (NFCR)—a leading cancer research charitable organization in the United States that has gained international recognition for its vision and unique approaches to accelerating cancer research toward a cure. The Prize is awarded annually to a scientist, nominated by colleagues or peers, who has contributed outstanding, significant research to the fight against cancer, and whose accomplishments have helped improve treatment options for cancer patients.The Prize is named in honor of NFCR co-founder Albert Szent-Györgyi, M.D., Ph.D., who won the Nobel Prize for Science and Medicine in 1937 for his discovery of vitamin C. The Prize and the surrounding ceremony also serve to promote public awareness of the importance of basic cancer research and to encourage the sustained investment needed to accelerate the translation of these research discoveries into new cancer treatments.Since its establishment in 2006, the Szent-Györgyi Prize has been awarded to 11 outstanding cancer researchers from around the world. Among the past awardees is Dr. Zhu Chen, chairman of the Chinese Medical Association and former Minister of Health of China, who won the 2012 Prize together with his mentor, Dr. Zhen-Yi Wang.In 2015, the Szent-Györgyi Prize for Progress in Cancer Research was awarded to Frederick Alt, Ph.D., Professor of Genetics at Harvard Medical School, Director of the Program in Cellular and Molecular Medicine at Boston Children’s Hospital, and Howard Hughes Medical Institute Investigator  at Boston Children’s Hospital. Dr. Alt has been Professor of Genetics at Harvard Medical School since 1991 and Charles A. Janeway Professor of Pediatrics at Boston Children’s Hospital since 1993. Prior to 1991, Dr. Alt was on the Faculty at Columbia University. He earned his Ph.D. with distinction from the Department of Biological Sciences at Stanford University in 1977. Dr. Alt is a member of the National Academy of Sciences, the Institute of Medicine of the National Academies, and a fellow of the American Academy of Arts and Sciences, the American Academy of Microbiology, and the American Association for the Advancement of Science. He is also an investigator at the Howard Hughes Medical Institute.Dr. Alt has won numerous honors for biomedical research, including the Stanford University Medical Center Arthur Kornberg and Paul Berg Lifetime Achievement Award in Biomedical Science and the National Cancer Institute’s Alfred Knudson Award for “pioneering contributions that have revolutionized the field of cancer genetics.” Each year, the Cancer Research Institute of New York presents the Frederick W. Alt Award for New Discoveries in Cancer Immunology in his honor. Dr. Alt’s impact extends far beyond his own laboratory, for not only is he an outstanding researcher; he is also an exemplary teacher. In 2003, he received the American Association of Immunologists Excellence in Mentoring Award. Dr. Alt has mentored over 100 students and research fellows, many of whom have become leaders in immunology, genetics, or cancer biology.For Dr. Alt, cancer research has always been a personal endeavor. During his remarks, Dr. Alt spoke of how cancer claimed the lives of both his parents when he was a young child Fig. . “It didDrug resistance was a well-known problem in cancer treatment when Dr. Alt’s career began. His early research sought to understand how cancer cells become resistant to the drug methotrexate, and it was animated by the “somewhat heretical notion that cancer cells might be able to increase their numbers of gene copies.” At that time, in the early 1970s, conventional wisdom held that each person had exactly two copies of each gene—one copy from their mother and the other from their father—and that their genome was otherwise fixed and unchangeable.After building the technology to probe cells for individual genes, Dr. Alt applied this technology to cancer cells that had been treated with methotrexate. “We found that cells that are very resistant have hundreds or thousands of copies of the dihydrofolate reductase gene which confers resistance,” said Dr. Alt. “When we took the cells out of the drug, they actually lost the gene copies, really showing that mammalian cancer cells could have unstable genomes. They could gain genes when they were put under selection for drug resistance, and they could lose them when it was taken away. That was really a shocking result.”Dr. Alt’s discovery of gene amplification in chemotherapy-resistant cancer cells served to overthrow the old concept of a fixed genome, suggesting instead that cancer cells could change their genes to evolve resistance to treatments like methotrexate.The story became even more interesting when the same pattern of amplified genes was discovered in the childhood cancer neuroblastoma. “We knew from our methotrexate resistance research what amplified genes looked like under a microscope,” said Dr. Alt, but it was not clear which gene had been amplified in these tumors. It was at this time that Dr. Alt made the connection between gene amplification and carcinogenesis. “The idea we had was that there would be a gene that actually caused the cancer—not one that would cause drug resistance, but one that made the cancer grow better,” he said. This idea built directly off the recent discovery of viral oncogenes by previous Szent-Györgyi Prize winner Dr. Peter Vogt.myc, so they named it neuroblastoma-myc or n-myc. “That was extremely satisfying for us,” said Dr. Alt, “because it showed that for gene amplification it was not only a mechanism by which cancer cells become drug-resistant but also a mechanism by which they get generated.” Today, genomic instability is recognized as one of the hallmarks of cancer.Dr. Alt and his colleagues identified that the amplified gene in neuroblastoma was related to the viral oncogene Equally important to cancer research has been Dr. Alt’s work on the critical DNA repair mechanism called “non-homologous end joining” (NHEJ). Dr. Alt not only made the initial experimental findings that led to the discovery of this pathway but also carried out an ingenious series of experiments over many years in his lab in Boston, taking it apart piece by piece to understand how it works. This work linked NHEJ to protecting against a specific type of DNA damage called translocation, which is a major component of many cancers, especially leukemia and lymphoma.Both amplified genes and translocated genes are key components of the Precision Medicine paradigm, which is at the heart of 21st century medicine. By identifying the source of genetic abnormalities that drive both cancer development and drug resistance, Dr. Alt’s insights helped to revolutionize cancer diagnostics and treatment. His discoveries led to a wholly new approach to treating cancer—identifying these genetic abnormalities, and then selecting new drugs that target each specifically.In his closing remarks at the award ceremony, Dr. Alt said “Now is the time. We have the opportunity, and if we get the support I’m confident that basic cancer-driven research will help exploit this current technological explosion, [producing] more and more breakthroughs at an ever-increasing pace.” Through events like the Szent-Györgyi Prize, NFCR is working towards this goal, by raising public awareness of research and encouraging sustained investment in the future of basic science.The 2015 Szent-Györgyi Prize Selection Committee was chaired by James Allison, Ph.D., and co-chaired by Sujuan Ba, Ph.D. Other selection committee members included leaders in cancer research and drug development from academic institutes and biotech and pharmaceutical industries: Webster K. Cavenee, Ph.D., Ludwig Institute for Cancer Research; Carlo M. Croce, M.D., The Ohio State University; George D. Demetri, M.D., Dana-Farber Cancer Institute; Richard Gaynor, M.D., Eli Lilly and Company; Thomas J. Kelly, M.D., Ph.D., Memorial Sloan-Kettering Cancer Center; Mary-Claire King, Ph.D., University of Washington School of Medicine; Scott M. Lippman, M.D., Moores Cancer Center; Tak W. Mak, Ph.D., Ontario Cancer Institute; Alex Matter, M.D., Experimental Therapeutics Center & D3, A*STAR Singapore; Scott D. Patterson, Ph.D., Amgen, Inc.; Philip Tsichlis, M.D., Tufts University School of Medicine; Peter K. Vogt, Ph.D., The Scripps Research Institute; Owen N. Witte, M.D., David Geffen School of Medicine, University of California, Los Angeles; Qimin Zhan, M.D., State Key Laboratory of Molecular Oncology, China; and General Secretary Yi Michael Want, M.D., Ph.D., NFCR.2014—James Allison, Ph.D., Chairman of the Immunology Department and Executive Director of the Immunotherapy Platform for the “Moon Shots” Program, University of Texas MD Anderson Cancer Center, Houston, TX, US.2013—Alex Matter, M.D., Chief Executive Officer, Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore.2012—Zhu Chen, M.D., Ph.D., Professor, School of Medicine of Shanghai Jiao Tong University; Chairman, Chinese Medical Association; former Minister of Health of China, Shanghai, P.R. China.2012—Zhen-Yi Wang, M.D., Professor, School of Medicine of Shanghai Jiao Tong University; Honorary Director, Shanghai Institute of Hematology, Shanghai, P.R. China.2011—Beatrice Mintz, Ph.D., Professor and Jack Schultz Chair in Basic Science, Fox Chase Cancer Center, Philadelphia, PA, US.2010—Peter K. Vogt, Ph.D., Professor, Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, US.2009—Ronald A. DePinho, M.D., President, University of Texas MD Anderson Cancer Center, Houston, TX, US.2008—Carlo M. Croce, M.D., Director, Human Cancer Genetics Program; Director, Institute of Genetics, The Ohio State University, Columbus, OH, US.2007—Webster K. Cavenee, Ph.D., Director, Ludwig Institute for Cancer Research, San Diego Branch; Distinguished Professor, University of California, San Diego, CA, US.2006—Harold F. Dvorak, M.D., Mallinckrodt Professor Emeritus of Pathology, Harvard Medical School; Chief, Department of Pathology, Beth Israel Deaconess Medical Center, Boston, MA, US.


In the Funding section, two grant numbers from the funder National Natural Science Foundation of China are missing. The correct grant numbers: No.81372426, No.81372906, No.81202128 and No.81272974.


In Eurasia, studies of fluctuations in the borders of areas occupied by sibling species of this complex, as well as their relative proportions in the areas where they are sympatric are epidemiologically important.The burden of malaria infection in the modern world remains significant. Specific changes in the relative proportions of malaria vector mosquitoes, Maculipennis Complex species, in the south of Western Siberia over the past 25 years of the 20Anopheles mosquitoes collected over the period from 1973 to 2012. A total of 37 Anopheles samples  from the Ukraine, European Russia and the Urals were studied. To identify An. messeae s.l. cryptic species A and B, polymerase chain reaction and restriction fragment length polymorphisms of the second internal transcribed spacer rRNA genes sequences (ITS2 PCR-RFLP) were used.Species identity and chromosomal polymorphisms within each population were defined by cytogenetic analysis of polytene chromosomes of third- and fourth-instar larvae and adult females of An. maculipennis s.s. is expanding to the northeast at a speed of approximately 30 km per year. In 2008 or 2009, this species appeared in the Southern Urals. The emergence of An. maculipennis in this region was accompanied by a decrease in the proportions of An. messeae A and An. beklemishevi and by an increase in the proportion of An. messeae B within An. messeae s.l. It is highly likely that the southwestern border of An. beklemishevi distribution area could shift in the same direction as expanding area of An. maculipennis.An. maculipennis s.s. to the northeast, followed by changes in the species composition of Anopheles as well as their relative proportions and fluctuations in the species areas, exacerbate the epidemiology of malaria infection in Eurasia.The geographical distribution of the Palaearctic mosquito species of the Maculipennis Complex is undergoing a gradual shift. Changes detected in the species distribution can be considered as a component of the biocenotic process triggered by global warming. Both the warming itself and consequent expansion of  Anopheles family differ in their physiology and ecology, including temperature sensitivity. Thus, changes in regional temperatures can influence the geographical distribution of Anopheles species as well as their interactions in the regions where the species are sympatric. Due to variable abilities of different species for transmission of malaria [Plasmodium transmitters [Anopheles maculipennis Mg. taxon [Anopheles maculipennis s.s. distribution area, will provide important insights into the role of global climate change in geographical distribution of the Palaearctic mosquito species of the Maculipennis Complex and its disease-relevant consequences.Malaria remains a serious health issue in the contemporary world, being one of the major causes of death from infectious diseases worldwide. Global epidemiological situation in relation to malaria spread remains complicated , 2. The  malaria –9, globasmitters  to invadg. taxon , 10–12 wg. taxon –16. In lg. taxon , 18; in g. taxon –22. Diveg. taxon –25. We bAn. messeae taxon presents a major obstacle in understanding of speciation-related events within Palaearctic Maculipennis Complex. Based on non-random combinations of chromosome inversions in natural populations as well as assortative mating and different ecological features, as early as in 1984 it was suggested that An. messeae includes two cryptic species. These were provisionally named An. messeae A and An. messeae B [An. messeae as a single polytypic, polymorphic species. Another research group isolated a new species, An. daciae, from An. messeae[An. daciae appeared to be identical to An. messeae A [An. messeae cryptic species will be referred to as An. messeae A. An. messeae s.l. will therefore refer to both cryptic species A and B.Lack of consensus in views on the structure of esseae B . The exiesseae B . Some reesseae B , 28 stil messeae. Based oesseae A . For thiAnopheles larvae and adult females were used. Locations, time of collection and relevant numerical data are given in Table An. messeae karyotypes were performed using polytene chromosomes maps described in [An. messeae species A and B have almost completely overlapping inversion polymorphisms and can be identified by inversions on sex chromosomes; they also differ quantitatively, by the frequencies of autosomal inversions [An. messeae A. The variants 1L/1L1, 1L1/1L1 and 1L1/¬ almost never occur in the karyotype of An. messeae B. 2R/2R1 heterozygotes and 2R1/2R1 homozygotes almost always belong to An. messeae B [An. messeae s.l. populations are summarized in Table An. messeae A in the populations studied. While the frequencies of homozygotes 2R1/2R1 and heterozygotes 2R/2R1  allow us to estimate the proportions of An. messeae B. Thus, analysis of polytene chromosomes allows species identification only for those specimens which possess marker combinations of chromosome variants in their karyotype. An. messeae species A and B can be accurately identified by the taxonoprint analysis [BstF5I restriction enzyme [An. messeae s.l. specimens collected in Miass in 2012 were established by ITS2 PCR-RFLP.Third and fourth instar ribed in , 33. Detribed in . The invversions , 27. If esseae B , 27. Then enzyme , 35. TheAn. maculipennis in samples collected at approximately the same time increased from the north and east to the south and west, and ranged from 0% in the Middle Urals to 80% in the North Caucasus and the Ukraine.The proportion of An. beklemishevi was identified in the samples collected mostly in the northern and eastern regions, and its peak frequency was detected at the extreme northeastern location .An. maculipennis and An. messeae in Yablonovsky and Khadyzhensk, Murom and Verbovsky); they also can fluctuate significantly during the same breeding season and even over several days within the same location and biotope .The proportions of sibling species may differ in samples collected simultaneously in closely situated locations, yet from different biotopes , An. maculipennis, An. messeae s.l. and An. beklemishevi may reside in the same biotope, as may An. maculipennis and An. messeae s.l. or An. messeae s.l. and An. beklemishevi; however, An. maculipennis and An. beklemishevi do not occur together in the absence of An. messeae s.l.Analysis of species composition from different regions and years Table  revealedAn. messeae s.l. is the widest among species studied. Borders of areas occupied by An. maculipennis and An. beklemishevi most likely reflect their responses to abiotic factors, which are commonly changing geographically. Furthermore, competition between the latter two species during larval development is higher compared to competition of each of them with An. messeae s.l.Elements of these observations are in agreement and show ecological diversification of the species at different stages of individual development. In particular, combinations of co-inhabiting species indicate that the ecological niche of An. maculipennis in both zones, with a significantly higher proportion in the area with a higher population density (p < 0.05). This could be interpreted as a result of species-specificity in egg-laying sites of An. maculipennis that can give rise to microlocalities with higher density of this particular species. Another plausible scenario is active larval migration in accordance with ecological preferences. In either case, the specificity of distribution is due to the behavioral characteristics. The key factor here is how behavioral reactions of each species reflect its competing abilities. It was experimentally shown that, at least under laboratory conditions, if maintained together at a high density, larval survival of An. maculipennis is superior to that of both An. messeae s.l. and An. beklemishevi[An. maculipennis and An. messeae s.l. larvae to water composition (both are found in the brackish water of Lake Muldakkul), An. maculipennis can outcompete under optimal combination of other abiotic factors. However, the species that loses in the larval competition is still able to use atypical habitats with extreme conditions. Thus, in the waters of drainage canals from rice fields in Yablonovsky village , characterized by high saprobity and a high density of amphibians, only An. messeae s.l. larvae, although at an extremely low density, were found. In addition, abiotic factors, which can compensate for inferior survival of a species at the larval stage by increasing its survival at the imago stage, may favor species coexistence in certain territories. An increase in the proportion of An. maculipennis in the southwest and south, generally coinciding with the isotherms of the warm time of the year and shorter periods of cold weather, indicates the primary role of temperature (through both direct and indirect effects) in determining the outcome of interactions between the species. This is in agreement with an idea proposed by Ushakov [An. maculipennis females start leaving their wintering eustatic shelters at an average daily temperature of 7.1°C [An. messeae s.l. this index was 4–5.0°C [An. maculipennis females, delay the start of reproduction phase and apparently shift generations of species relative to each other. In the samples, this shift would manifest as an increase in the proportion of one of the species.Analysis of mosquito larvae collected in different parts of the Pshish river backwaters, spaced by 5–6 m and characterized by variable population density , this species was found in a number of ecologically distinct habitats and in different years, therefore, its presence in Miass and consequently in the South Urals is now confirmed. The study of samples collected in Lake Muldakkul and in Koltashi village in 2012 revealed the presence of An. maculipennis in the first location and absence in the second, where An. beklemishevi was found.In line with temperature trends in northern Eurasia over the last 40 years, the distribution area of  in 1989 . Howeverapproach , the speAn. maculipennis was first identified in 2010 with a frequency similar to that reported for the northeast periphery of this species area in 1986–1987. Its low frequency in different biotopes suggests that it most likely emerged in Miass in 2008 or 2009. This allows estimating the rate of An. maculipennis expansion to the east.Of note, in Miass, An. maculipennis appeared in Miass are separated by 22 years and about 750 km, and assuming that the border of the distribution area at that time was already shifted to the east of Samara, the average speed of the border movement eastwards is about 30 km per year. If true, this speed is significantly higher than estimated for other insect species, − 16.9 km per decade [An. maculipennis is an endophilic species [An. maculipennis, and the vector of its expansion is from urban areas to the countryside. Taking into account the key role of temperature in the life cycle of mosquitoes and isothermal patterns in the European part of Russia and the Urals, the northern border of the An. maculipennis habitat is likely to move at the same speed, about 600–700 km further to the north of the border reported in 1986–1987 [et al. (2012) [An. maculipennis s.s. in Karelia up to the town of Kem’ . However, the distance between Rostov, where An. maculipennis at low frequency was discovered in 1986, and Kem’ is over 1000 km. In the literature, there is also a discrepancy in the description of the species composition in Kem’, where, according to Moskaev [et al.[An. maculipennis and An. beklemishevi. Neither Stegnii et al.[An. maculipennis and An. beklemishevi in the absence of An. messeae s.l. in any of the geographical locations studied. Based on the data available so far, it can be assumed that the probable northern border of An. maculipennis habitat lies at a latitude of 64°N, i.e. to the north of the town of Segezha.Given that the time of sample collection in the eastern locality for the earlier study  and the most probable time when r decade . Because species , 42, its986–1987 . Moskaev. (2012)  found An [et al. found oni et al., nor MosAn. maculipennis to the north and east is a part of the biocenotic process caused by global warming. Changes in the borders of distribution areas and/or proportions of sibling species in the zones where the species are sympatric are also components of this process. The rate of climate warming has been estimated at 0.5°C per 100 years in the northern parts of European Russia and 1.4-1.6°C per 100 years in the south of the Urals, with the largest incremental changes in the past several decades [An. maculipennis is given a better opportunity to invade the regions adjacent to the northern and eastern borders of its current distribution area. Expansion of An. maculipennis to the northeast is accompanied by a decrease in An. messeae s.l. and An. beklemishevi frequencies and probably even disappearance of the latter species. An. beklemishevi was found in the surroundings of Priozersk, Syktyvkar and Chelyabinsk in 1975 [An. beklemishevi could be absent due to environmental and behavioral characteristics of this species. Moskaev (2012) [An. beklemishevi in Priozersk and confirmed its lower frequency in Syktyvkar in 2010 compared to the report of Stegnii et al. in 1978 [An. beklemishevi in Zelenodolsk coincided with the emergence of An. maculipennis[An. maculipennis, this species will expand to the east within borders very similar to that of An. messeae A [An. maculipennis and both cryptic species of An. messeae s.l., as well as increasing contacts between An. maculipennis and An. beklemishevi. At the larval stage, under conditions of high-density, An. maculipennis demonstrates superior survival compared to An. messeae s.l., i.e. both A and B species [An. messeae A and An. maculipennis are considered as endophilic species, while An. messeae B is exophilic [An. maculipennis and An. messeae A have very similar environmental needs, and their ecological niches largely overlap, leading to their competition and as a result, reduction in the frequency of the latter species. Indeed, cytogenetic analysis showed that An. maculipennis emergence in Miass was accompanied by a change in the proportions of the An. messeae cryptic species. In line with this finding, we detected a highly significant (p < 0.001) decrease in the frequencies of homozygotes, heterozygotes and hemizygotes for inversion 1L1, as well as other cytogenetic markers of An. messeae A, over the period from 1999 to 2011, accompanied by a corresponding increase in the frequencies of markers typical for the An. messeae B , for these the presence of the inversions on X-chromosome was also determined. Only one male, identified by ITS2 PCR-RFLP as belonging to An. messeae B, had variant 1L1/¬. Thus, the inversion 1L1 is highly specific for An. messeae A  from earlier to later samples, as defined based on marker inversion variants, reflects the actual sequence of events. This allows us to estimate the proportions of An. messeae cryptic species A and B by analyzing the frequencies of the above cytogenetic markers in other regions and locations. Analysis of species proportions in different geographic locations   did not  in 1978 . Previoulipennis. Steady esseae A , i.e. as species . An. mes12  did nLP Table . Speciesoven yet ). Thus, An. maculipennis is high or it is dominant over other species, the frequency of the cytogenetic markers typical for An. messeae B is relatively high. However, in areas where An. maculipennis is absent or rare, the proportion of An. messeae A is increased, which is signaled by high frequencies of chromosome variants typical for this species . In some regions this correlation is not observed (Yoshkar-Ola), which is likely due to the low numbers of An. messeae A individuals in this region. In Ciscaucasia and in the suburbs of Kharkov, in accordance with its cytogenetic structure, only An. messeae A was found in conjunction with An. maculipennis. In Khadyzhensk and in the suburbs of Kharkov, the latter species was predominant  of An. maculipennis. It is also possible that the observed trends in species frequencies result from not only competition between An. maculipennis s.l. species, but also from alterations in other ecosystem components. Our results describing the impact of An. maculipennis on the ratio of An. messeae species A and B are consistent with the situation described in Germany, where An. messeae s.l. and An. maculipennis are sympatric. According to the results of sequencing of marker portions of genomic DNA from mosquitoes collected in localities with low (a few percent) and high (over 50%) occurrence of An. maculipennis, An. messeae species referred here as the species B, outnumbered the other one [For example, in Solnechnogorsk, Suzdal, and Protvino where the proportion of nt Table . Despitether one .An. maculipennis expansion to the Urals, and likely, soon after that, to Western Siberia can be foreseen. In parallel with a decrease in the number of An. messeae s.l., the proportion of An. messeae A cryptic species will also decline, together with a respective increase for An. messeae B. The latter will manifest itself through increasing karyotype variability, i.e. a process opposite of that previously described [An. maculipennis will follow the path of colonization of Siberia pursued earlier by An. messeae A, which is characterized by similar environmental needs but is somewhat more resistant to low temperatures. Expansion of An. maculipennis to the north and east will continue until an equilibrium is achieved among the four species, two of which (An. maculipennis and An. messeae A) are endophilic and thermophilic and the other two (An. messeae B and An. beklemishevi) are exophilic and cold-resistant. This balance will be maintained mainly due to different durations of the reproductive period typical for each species, during which a growth in the frequencies of the first two species is expected, and this growth will be more pronounced for An. maculipennis. The second factor to play a role will be the availability of wintertime eustatic shelters and temperature in those during the cold time of the year, when the numbers of the first two species will decline more rapidly. These conditions will be less severe for a more cold-resistant An. messeae A compared to An. maculipennis. Expansion of An. maculipennis is likely to be accompanied by ousting of An. beklemishevi from more or less populated areas, and correspondingly, the southern and western borders of the distribution area of the latter species will move to the north and east. However, because An. beklemishevi is exophilic, it will still be able to survive, although in small quantities, in the wild. Given that An. maculipennis is able to transfer Plasmodium vivax, and that global warming leads to increased species diversity and abundance of Anopheles on large territories, one can confirm a worsening epidemiological situation in Eurasia.Possible consequences of escribed . Thus, AAn. maculipennis geographic and temporal distribution made it possible to reconstruct the dynamics of the area occupied by the species  whose dynamics would significantly modify the effect of all other factors. In the northeastern periphery of the An. maculipennis distribution area, sympatry with An. messeae s.l. is compulsory, with An. beklemishevi – possible; coexistence of An. maculipennis and An. beklemishevi in larval habitats in the absence of An. messeae s.l. is an exception. On this territory, one can expect cyclic fluctuations both in the number and the frequency of An. maculipennis, with ups during the warm (reproductive) season and downs during wintertime.Significant increase in the global temperature, i.e. global warming, recorded in the past 40 years, has resulted in the changes of Maculipennis Complex species distribution in Eurasia. The distribution area of


World Water Day, sponsored by the United Nations, is observed each year on March 22. This year, World Water Day focuses on water and energy. Water and energy are closely linked. Worldwide, 1.3 billion persons currently live without electricity (To some degree, water is crucial to produce, transport, and use all forms of energy, and these activities variously affect water resources. Demand for freshwater and energy will continue to increase significantly over the coming decades, presenting major challenges and straining resources in nearly all regions. This is especially true in developing and emerging economies. Better understanding of the linkages between the water and energy sectors can lead to improved coordination among policymakers, planners, and others in providing and obtaining more efficient and cost-effective water and energy services.http://www.unwater.org/worldwaterday. Information on CDC’s efforts to ensure global access to improved water, sanitation, and hygiene resources is available at http://www.cdc.gov/healthywater/global.Additional information about World Water Day and ideas on how to get involved are available at


Nigel Kiely.In the original publication, the name of one of the co-authors has been published incorrectly. The correct name should be Furthermore, the affiliation of Dr. Rohit Singh and Dr. Stuart Hay is included now.Royal Shrewsbury Hospital, Shrewsbury, Shropshire SY38XQ, UK.


The grant number in the funding statement is incorrect. Please refer to the correct funding statement below:www.fwf.ac.at <http://www.fwf.ac.at>). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.This work was supported by a grant from the Austrian Science Fund (Project P20.196-B17) (


Fagus sylvatica L.) and Scots pine (Pinus sylvestris L.) was intra-annually monitored to examine plastic responses of the xylem phenology according to altitude in one of the southernmost areas of their distribution range, i.e., in the Moncayo Natural Park, Spain. The monitoring was done from 2011 to 2013 at 1180 and 1580 m a.s.l., corresponding to the lower and upper limits of European beech forest in this region. Microcores containing phloem, cambium and xylem were collected biweekly from twenty-four trees from the beginning of March to the end of November to assess the different phases of wood formation. The samples were prepared for light microscopy to observe the following phenological phases: onset and end of cell production, onset and end of secondary wall formation in xylem cells and onset of cell maturation. The temporal dynamics of wood formation widely differed among years, altitudes and tree species. For Fagus sylvatica, the onset of cambial activity varied between the first week of May and the third week of June. Cambial activity then slowed down and stopped in summer, resulting in a length of growing season of 48–75 days. In contrast, the growing season for P. sylvestris started earlier and cambium remained active in autumn, leading to a period of activity varying from 139-170 days. The intra-annual wood-formation pattern is site and species-specific. Comparison with other studies shows a clear latitudinal trend in the duration of wood formation, positive for Fagus sylvatica and negative for P. sylvestris.Wood formation in European beech ( A forest community can prosper only on sites where the environmental conditions are within the niche volumes of each species . The disFagus sylvatica) and Scots pine (Pinus sylvestris) have both high ecological relevance and economic values. European beech forests are spread all over central Europe, from central Poland, the south of Scandinavia and British Isles to the southernmost locations in the mountain ranges of Greece, Italy and Spain  and the timing of xylem formation was compared between the two species and sites to evaluate the adaptation strategies under different environmental conditions. The duration of the xylogenesis was compared with data from other studies of the same two species performed all over Europe.In order to better understand the growth adaptations and limitations of F. sylvatica and P. sylvestris. The mean annual temperature and average annual precipitation for the last 37 years were 11° and 710 mm, respectively .F. sylvatica forest on this mountain. The low elevation site was located at 1180 m a.s.l. and the high elevation site at 1560 m a.s.l.Two pure stands of different altitudes were selected for each species, corresponding to the lower and higher altitudinal limits of P. sylvestris and 35–50 cm and 120 years for F. sylvatica. From each tree, two microcores containing phloem, cambium and the last formed xylem growth ring were collected at breast height with a Trephor tool , cells undergoing secondary wall thickening and mature cells. For P. sylvestris, the cambial cells were counted as well as the xylem cells in the three different aforementioned phases.For Cambial activity was identified and interpreted within the context of the multi-seriate concept, that the vascular cambium comprises both the cambial initial cells and xylem and phloem mother cells . Thin-waP. sylvestris to normalize the measurements according to F. sylvatica, the normalization formula was adapted using width measurements instead of cell number, as described in The number of cells and the width of tissues in each phase varied between and within trees due to the variation of the tree-ring width around the tree circumference. The number of cells in the previous xylem ring was therefore counted for Extreme values were filtered and xylem-formation dynamics was analyzed with the Gompertz function . CambiumFigure 1).In all years, locations and species, the cambium was still dormant on the first sampling date in the last week of March. Despite site and annual variations in weather conditions, the different xylogenesis phases followed a common pattern during the growing season. The cell enlarging and cell-wall thickening curves follow a characteristic bell shape, while the cell maturation curve follows a sigmoid shape . The cell enlargement started first in P. sylvestris, around 31 March (DOY 90) and 50 days later, around 20 May (DOY 140) in F. sylvatica. Cessation of cell enlargement was observed between 9 July and 6 August (DOY 190-218) for F. sylvatica and in P. sylvestris between 10 August and 1 October (DOY 222-274). Cell-wall thickening and lignification began up to ca. one month earlier and ended around three months later in P. sylvestris than in F. sylvatica. The first mature cells were observed around 25 June (DOY 176) in P. sylvestris and around 7 July (DOY 188) in F. sylvatica.The critical dates for the xylogenesis of ∗site: p < 0.001). In F. sylvatica, it began between 7 May and 20 June (DOY 127–171), with noticeable differences within years . In P. sylvestris, cell enlargement began between 23 March and 17 April (DOY 82–107). Although the variability was lower in the latter species, in 2011 there was a delay in the beginning of cell enlargement. Focusing on the elevation differences, cell enlargement started earlier at lower elevation in the P. sylvestris than in F. sylvatica. The end of this phase highly varied among years, whereby higher variability was observed between the years then among the sites for each species (ANOVA site: p = 0.31).The beginning of the enlargement phase was highly variable and significantly different among the years for both species at both high and low elevations ; in all cases the cell-wall thickening phase started earlier in 2011 and later in 2013, followed by the same temporal pattern (within-subjects ANOVA time∗species∗site: p = 0.561) . The first mature cells (5%) were formed earliest in P. sylvestris at low elevation in 2012 , and in 2011  for F. sylvatica.The onset of the secondary wall formation was highly variable (F. sylvatica was significantly shorter than in P. sylvestris (ANOVA species: p < 0.001) . The cell production period during the three study years took 48–75 days for F. sylvatica, in contrast to P. sylvestris, with a growing period lasting from 140 to 170 days. Trees growing at low elevation had a longer growing period in the case of P. sylvestris, whereas the growing period of F. sylvatica, in contrast, was shorter at low elevation than at high elevation.The total duration of the xylogenesis of Table 1 and Figure 4). In P. sylvestris, the duration of xylogenesis was shorter at high latitude and longer at low latitude, with a range of 49 days in Finland .The mean duration of the xylogenesis was compared with other studies all over Europe ( Finland  to 217 dherlands  and onlyF. sylvatica and P. sylvestris respond differently to local Mediterranean conditions. Accordingly, the phenology of xylem formation was significantly different between the two species, the period of all P. sylvestris developmental phases being significantly longer.Tree growth is largely affected by different climatic conditions, which become more limiting in adverse climatic conditions, such as in a Continental Mediterranean climate . DiffereP. sylvestris than on F. sylvatica, despite this early-successional condition. The Pinus genus has been established as very plastic and capable of adapting its growth to changing climatic conditions (P. sylvestris has recently been determined as a plastic species in the Mediterranean area (Our results demonstrate that Mediterranean climate has less impact on nditions  and the nditions . Specifiean area .F. sylvatica under Mediterranean conditions have highlighted the growth limitation due to summer high temperatures and drought (F. sylvatica trees are more limited during the year in terms of plasticity because, with the activation of a leaf senescence mechanism, trees inexorably enter a dormant period. Despite this, our results reveal differences in the altitudinal gradient in agreement with the results shown in F. sylvatica forests with different climatic regimes.Several studies performed on  drought . In addiF. sylvatica, similarly as in P. sylvestris.The most striking result of the present study is the great differences in growth patterns among the years, highlighting a plastic response of radial growth in High variability in xylem phenology between years and sites demonstrates high plasticity of the species. The timing of different developmental phases significantly varied between the two species. Even though the variability of the critical dates was high among years and sites, the most remarkable disparity was found between the two tree species.F. sylvatica started earlier than in the other two study years, while P. sylvestris showed the latest onset of growth in the same year.Xylogenesis, starting with cambial division and cell enlargement, is triggered by an increase in air temperature in spring. Several studies have demonstrated this positive relationship , which hP. sylvestris continued until mid-November, whereas in F. sylvatica it ceased in mid-August, i.e., about three months earlier. Mild temperatures during early autumn may result in an extension of the growing period for P. sylvestris but not for F. sylvatica, since by that time leaf senescence has also already started. Cessation of cell-wall thickening was first observed in the lower part of the mountain in both species, as was similarly reported by The end of the cell-wall thickening phase seems to be a key date, since it defines the end of xylogenesis. Overall, the thickening phase in P. sylvestris. This may be explained by the late frost that occurred in 2013, after the onset of cell enlargement in P. sylvestris. This event presumably affected the temporal dynamics of wood formation in both species, although with different magnitudes. F. sylvatica. Nonetheless, a more detailed study of the climatic-growth relationship would be needed to confirm this hypothesis.A common pattern is an evident delay in all developmental phases in 2013, except the beginning of wood formation in F. sylvatica and more than five months for P. sylvestris. Because of the early start and late end, P. sylvestris on Moncayo showed the longest xylogenesis duration in this species recorded in the various studies to date. Moreover, the shortest xylogenesis duration for F. sylvatica was also captured on this mountain.Although both species are growing at their southern distribution limit and, consequently, their radial growth is somewhat constrained, the duration of the growing period of the two species significantly varied. The duration of xylogenesis highlights the differences between the species; it was two months for P. sylvestris occurred earlier than on numerous Central European sites (P. halepensis (P. pinaster (F. sylvatica, the results showed the opposite response, the beginning was later than in colder locations in Europe, such as in France (F. sylvatica.Cambial resumption in an sites , which clepensis  or P. pipinaster . The extn France , Slovenin France , Romanian France , Czech Rn France  and the n France . This inF. sylvatica than in P. sylvestris growing at the edge of their southern spatial distribution. This shows that intra-annual radial growth patterns in the studied species are differently affected by the Mediterranean conditions. The annual variation of the critical xylogenesis dates indicates a high species-specific plasticity for adapting to changing climatic conditions. As a result, the period of xylogenesis in F. sylvatica was around 2 months, while for P. sylvestris it was more than 5 months. Our findings are in accordance with our hypothesis of contrasting growth strategies and adaptations of the two species at the edge of their spatial distribution.It appears that the temporal dynamics of xylogenesis is considerably different in P. sylvestris showed a positive xylogenesis duration trend on southern locations. F. sylvatica, in contrast, showed a shorter xylogenesis duration in the south of Europe than that shown in northern locations. These findings demonstrate that a deciduous and late-successional species such as F. sylvatica is negatively affected by Mediterranean climatic conditions, resulting in a shorter xylogenesis, whereas in the evergreen and early-successional P. sylvestris, xylogenesis is shown to be longer in Mediterranean environments.Furthermore, we compared our observations with those of other authors working on the same two species in different climatic environments, especially along a latitudinal range. A clear north–south trend was found in the xylogenesis duration over the distribution range of both species. EMDC did the fieldwork, wrote the paper and together with ML carried out the statistical analysis and prepared the figures. ML and LA developed the idea of this work and get the necessary funds. EMDC together with KČ, JG, and PP did the laboratory work and measurements. EG-P and KČ provided facilities and material support and gave technical support during laboratory work. All authors without exception helped to improve the work, specially the discussion and conclusions.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.