Positive psychology has accumulated a large and ever-growing body of scientific knowledge about human strengths and virtues. However, research on positive psychology interventions (PPIs) to develop such is still in its infancy. In this brief position paper, we summarize the status of PPIs in one of the positive psychology’s most important virtues: temperance. Temperance refers to the capacity to manage habits and protect against excess and is composed of forgiveness, humility, and (we include) patience. Specifically, we examine the current state-of-the-science in the conceptualization of temperance, explore the efficacy of temperance interventions, and reflect upon what the future may hold in this research domain. In this paper, we first highlight the challenges and opportunities for expanding the theoretical conceptualization of temperance and reflect upon the challenges in temperance-related PPIs. For each aspect of temperance, we propose a specific research agenda. Second, we explore what is needed for PPIs to promote temperance and how growth in temperance intervention research can be fostered. Generally, while forgiveness interventions are well established, we recommended that both humility and patience interventions need more viable evidence-based research on existing and new interventions. Third, we advanced several recommendations regarding how to promote more research in new interventions. These recommendations included attracting more funders to the area, developing new interventions, and employing new technology. Because intervention research in temperance is in its infancy, the future looks rosy for PPI researchers as we move into a second generation of positive psychology research. Positive psychology has accumulated a large and ever-growing body of scientific knowledge about character strengths and virtues . HoweverEach virtue is important, and in many ways, all virtues are interconnected. However, some seem more fundamental than do others because they organize or empower others. For example, humility, which is not strictly a virtue in forgiveness and mercy, humility, prudence/self-regulation, and (we would add) patience  a transgressor (2) someone over whom one has power or authority or (3) someone in great distress”  and benevolent motivations  within an essay of reasonable length. We generally made our recommendations because (1) they had not been previously researched and (2) they seemed to us to be needed, given our subjective sense of what might be needed to push ahead research on each topic. We hope our suggestions are heuristic, and we apologize for the lack of detailed context around each one. With that caveat in mind, we believe the next decade in forgiveness research should as:Determine the growth-promoting and transcendence functions of forgiveness, neither of which have received much research given the focus on the deleterious effects of not forgiving characterizing much research ;Find differences in the ways that decisional and emotional forgiveness interact with each other over time and ways that each contribute to spiritual, relational, psychological, and physical health outcomes; whereas the two types of forgiveness have been hypothesized to have different spiritual, relational, psychological, and physical health outcomes, little systematic research has investigated what those differences are ;Enhance and supplement theoretical and empirical frameworks for self-forgiveness; Map interconnections, especially over time, among decisions to forgive, emotional forgiveness, observing in offenders expressions of accountability , and observing in offenders indications of self-forgiveness and feeling forgiven by God ;Explore context-specific variables that affect the presence or absence of forgiveness in dyadic, family, group, community, and macro  contexts  in this era of increasing political and ideological polarization that has resulted in a strong valuing of punitive justice of those who harm and censure of those failing to support social justice for those who lack social power .Humility is a character strength rooted in the work virtuous ethics. It is a critical component of relationships providing social oil that facilitates smooth relationships and a buffer against hostility and relationship-disintegration when things go wrong in relationships. Humility also directly affects mental health and wellbeing , and it realistic self-assessment through interactions with others. Humble individuals process information gathered in social interactions as a means to both explore their personal identities  and to modify their behavior when necessary. Humility, therefore, fosters an accurate or objective appraisal of one’s own strengths and areas needing additional development through a transparent (situation-appropriate) disclosure of their own limitations, openly acknowledging their mistakes, and actively seeking constructive feedback on their own development. Humble individuals tend to show high levels of relational functioning, psychological wellbeing, and mental health . Relational humility is humbleness that is differentially experienced in different relationships with others such that, for example, being a humble boss or worker would entail different manifestations. Cultural humility is humbleness about cultural beliefs, differences, and views. Intellectual humility is humility about personal opinions, ideas, and intellectual capabilities that one is emotionally invested in. Finally, spiritual humility is humility before what one considers sacred.Although humility has over 300 studies investigating it, it is still concept within positive psychology that requires further exploration. We therefore see a number of possibilities that could strengthen then scientific foundation of humility research. Future research on humility should focus on:Clarifying the definition; about half of the researchers see the definitional point of being other oriented to be debatable; and many favor “a quiet ego” as a necessity. Future research should resolve whether that point is affected by collectivistic versus individualist cultures or relationships, by religious beliefs . Such distinctions have not been addressed. While there might be religious and cultural origins to seeing humility as requiring a “quiet ego” or other orientation, it is unclear whether there is any residual religious basis for differing on this crucial point of the definition of humility;Articulating an evolutionary theory of humility’s benefits to the species. There are no evolutionary accounts of humility in the literature;Describing how humility (with its emphasis on lifting others up), when applied to disadvantaged minorities, is not a call to accept oppression, but rather is a call to social justice. This has long been a criticism of humility from theoreticians writing about implications for people of disempowered status , (2) life hardship patience , and (3) daily-hassles patience . Managing suffering at various levels from intense pain to aggravating unpleasantness is therefore a core component of patience. The magnitude and severity of the causes of the suffering to be endured are objectively different among these types of patience. However, all three types of patience involve an individual’s calm response to what is subjectively perceived as a source of minor or major suffering  needs be established for it to thrive as a stand-alone strength .Because calm waiting, rather than mere waiting, is hypothesized to be essential to patience, experimental determination of how calm one must be and how that is manifested physiologically, cognitively, and behaviorally need to be mapped  they act as conduits for victims to reflect upon and disclose the details of a transgression (2) they aid in altering or reframing the perspective through which transgressions are interpreted (3) they seek to develop empathy, kindness, and understanding for the transgressor, and (4) they promote intentional commitment to forgive transgressors  in standard deviations and the time spent conscientiously trying to forgive . This dose–response relationship has been described using a regression equation, d = 0.124 + 0.046*T. The initial intercept is likely a positive effect from hearing a definition of forgiveness and a summary of the relational, psychological, and physical health benefits of forgiveness, which have been rigorously accumulated from basic research on forgiveness. Forgiveness interventions have been beneficial for increasing experiences of forgiveness, hope, optimism about the future, happiness, and wellbeing —humility, forgiveness, patience, self-control, positivity, and no-action—the undergraduates who completed a DIY workbook on PROVE Humility (n = 26) outperformed a non-action control condition (n = 33). When participants were recruited specifically into a study to promote humility, the gains for the workbook (n = 39) relative to the control condition (n = 33) were about twice as big. Although the results of the interventions were promising, the studies were both with students at a single institution. n = 41) beyond a wait-list control condition (n = 30) in their study of religious leaders. For humility interventions, the PROVE Humility model , 2017 anHumility interventions appear more difficult for which to recruit participants than are forgiveness interventions. When people struggle with forgiveness, they feel a need. Grudges are unpleasant and revenge motives might frighten people, so they seek ways to eliminate those experiences and forgiving is one salient possibility. However, most people do not consider a deficit of humility to be a problem. In fact, one might be arrogant, and thus conclude that one does not need humility, already perfectly having that strength. One might exhibit too many self-effacing behaviors and feel oneself to be a doormat, but low self-worth might make the person feel unworthy of pursuing humility. Thus, the “market” for humility interventions might be within communities that value humility. These might be religious organizations. But some business organizations operate using a strong ethic of teamwork, based on other-oriented humility. Paradoxically, even armed forces, which train soldiers to be loyal to the small group, might benefit by humility interventions. Finally, Patience interventions have been studied more frequently than have humility interventions. Many interventions with adolescents to promote patience are brief and are aimed at stimulating better performance on frustrating tasks, like writing with the non-dominant hand . InterveN = 71; 61 female) experienced an end-of-treatment increase in trait patience and positive affect but not a decrease in negative affect and a decrease in depression. Gains in trait patience, however, were not maintained at follow-up.n = 28) produced more trait patience, trait self-control, and forgivingness than initially, relative to the control condition (n = 33).Yet many adults over 26 years old struggle with failures in patience. So, assuming maturation will infallibly occur by age 26 is not a fruitful strategy for many. We also must help mature adults to build up a habit of experiencing patience. The interventions created by Schnitker et al. and Despite significant advancements in establishing temperance as an important concept within the larger nomological network of psychology, and the magnitude of PPIs aimed to develop its core components, we believe that two core questions remain unanswered:What is needed for interventions to promote forgiveness, humility, and patience?How do we promote growth in temperance intervention research?We hope that evidence-based temperance PPIs can move beyond efficacy research to effectiveness research and from effectiveness research to scaled up dissemination methods by answering these two questions. We briefly reflect upon each of these questions in a hope to stimulate future research.Forgiveness interventions are well proven at the small, efficacy-study level . They reAnother factor to consider is the contextualization of forgiveness interventions. Forgiveness interventions should, therefore, not only focus on developing forgiveness within the full interpersonal context but also as applying in intergroup contexts as one means to enhance its global applicability . It is iFurthermore, even with evidence-based psychoeducation, psychotherapy, couple therapy, and prevention interventions, effectiveness studies are also needed. Rather than continuing to produce additional closely controlled, lab-based efficacy studies in cultures that have already been investigated, controlled effectiveness studies in real-world contexts are needed.Dissemination studies are also needed in which organizations adopt a particular approach to forgiveness and investigations must be conducted to monitor systems  to ensure fidelity of delivery and investigate the effects on the entire system . ScalingFurthermore, shifts in the demand for forgiveness interventions would affect both consumers and creators. Both will be looking to create or engage in new, brief, punchy, emotionally engaging, and motivating interventions that frequently engage the consumer in providing feedback and receiving level-up feedback to reward continued participation . This wiHumility and patience have struggled to attract intervention researchers—for different reasons. Humility has struggled because people do not experience as much subjective discomfort from evaluating themselves as not humble as in evaluating themselves as unforgiving. Humility indeed has social benefits that have been well documented see —providinFurthermore, limited empirical and theoretical models exist that explain the routes toward humility. Despite being a valued virtue, there is little empirical evidence for the factors contributing to or influencing the development of humility . WithoutDespite this limitation, at least one of the two interventions aimed at developing humility showed promise. Both interventions employed traditional self-administered activity designs (such as workbooks), which were aimed at self-exploration. These interventions are self-paced and do not require the presence of a facilitator/therapist. However, these interventions relied on DIY workbooks. Although they were completed on a computer and submitted online, they we essentially traditional. Electronic formats  are needed to supplement the DIY workbooks and facilitated interventions must be developed for application in local communities if humility interventions are to be scaled.Another element to consider is the paradoxical duality of humility. Humility is a personal function (or private experience) but can only exist (or be expressed) in the presence of others. Finally, humility interventions should be expanded beyond the mere focus of general humility. Humility intervention researchers should develop intellectual humility , spiritual humility , relational humility , and cultural humility interventions. Multi-facetted approaches toward humility development could therefore be greatly beneficial.Patience interventions have been more frequent than humility interventions yet are plagued with similar issues. Additionally, patience interventions have almost exclusively been relegated to children, adolescents, and early adult college students . For exaMoreover, despite the limited agreement on the components of patience, most interventions are exclusively focused on regulating behavior and emotional management. This has shown to be effective in certain population groups; however, the long-term sustainability thereof has been questioned . PatiencFinally, interventionists and creators of interventions should be conscious of the “consumer culture” in mental health-related practices. People who need to build their patience would ironically be saying, “I want patience—now!” Creators of PPIs to promote patience must be cognizant of their audience’s inherent bias against sticking with patience interventions patiently—unless the creators heed new strategies to promote engagement .One of the things that would increase the stable of researchers is additional grant funding. The turning point in research on forgiveness was the 1997 request for proposals that the John Templeton Foundation advanced. In the end, it funded about 20 research teams many of whom did multiple studies on forgiveness and that attracted post-doctoral researchers and graduate students who built their careers studying forgiveness. Money does not simply flow to a funding area. There must be a need and, following that, a funder whose priorities include building up the area. So what could create a need to study temperance?One potential need could occur is there was a crisis associated with intemperance could increase research in temperance. As we found in the COVID-19 pandemic, crises mobilize action  and can As we saw in 2020, injustices are widespread throughout the world. Many of those might be thought to have occurred because people with abundant resources have pursued self-interested goals to the expense of less privileged people. Humility on behalf of the privileged might be needed to reverse this trend by not assuming that they have entitlement to an unfair share of resources. Forgiveness is needed by people who have been on the short end of social injustices instead of revenge that can perpetuate injustices. Forgiveness happens internally, as decisions to treat offenders as valuable people lead to emotional change that replaces negative unforgiving emotions with positive other-oriented ones . But, foSome people believe that forgiveness by victims of social inequities might short-circuit the pursuit of social justice. However, forgiveness is not opposed to justice. In fact, justice is a social and societal phenomenon and forgiveness in internal, involving a decision to treat people less hatefully and more benevolently and an emotional transformation involving less rumination and thus less depression, anxiety, and anger. It is possible that victims’ forgiveness can remove people’s focus on themselves, or their expenditure of energy direct at rage and hatred, and thus leave them more able to pursue social justice. In addition, offenders and those who have the position to speak to historic wrongs from the offenders’ perspective need to be aware that their public statements of contrition, apology, and regret can encourage more forgiveness by victims, and amends making can begin or further reconciliation.The awareness of increasing political and social polarization could create a sense of need to pursue a widespread building of temperance. This might involve political humility, forgiveness of people on the other side of the political spectrum, and patience as people wait for change to occur. Movements like global warming and environmental concern have been polarizing. If both sides could see the potential for temperance, it might lead to positive steps toward a healthier environment. In addition, geographic, financial evolution might create another sense of need for more global temperance. In Asia, China has risen as a technological, industrial, and financial power, and Japan has continued to be strong. In the Middle East, the worldwide weakening of oil dependence on Arab nations is likely to portend a shift in global economics. There will be ample opportunity to forgive any Arab nations that might have been perceived as holding economic power over much of the rest of the world. In Europe, the weakening of the European Union with the withdrawal of Great Britain provides opportunities to forgive both within the EU and Great Britain and in countries who have dealt with them. In South America, the economic and social strengthening of the southern hemisphere is a global trend. Some interventions have developed to treat unforgiveness that has come about from civil wars and other South American conflicts. Narvaez and Foundation ES.PE.RE is a noted example. In Africa, the rise in religious consciousness toward Christianity and in North Africa, Islam, have created the potential for religious tensions. These add to tribal tensions that have been long experienced in the Great Lakes region of East Africa, including Rwanda, Burundi, Uganda, and other countries. In Central America, emigration and political stabilization create needs for temperance—forgiveness, humility, and patience. In the United States, political polarization, conflict, the cancel culture, and divisions economically, politically, racially, and religiously are fertile grounds for practicing the virtue of temperance.When people build temperance, they are not passively reacting to circumstances requiring restraint. Rather they are actively seeking to restrain negative knee-jerk reactions. Such restraint might take different forms. For example, people could tolerate changes. Tolerance involves restraining oneself from negative social responses, even though internally one might feel negative emotions. Forbearance involves restraining negative reactions for a positive motive—for the sake of group harmony. That positive motive can therefore mitigate the negative emotions of tolerance while yielding the same restraint of social wrongdoing as tolerance. Acceptance speaks to restraint of negative reaction and a passive internal emotional state. The position of acceptance might be actively arrived at yet there is more equanimity than with either forbearance or tolerance. Forgiving restrains negative reactions and also more actively puts the past in the past, yielding a different type of emotional equanimity than acceptance. Building forgiveness and having a patient stance toward seeing change come about socially, while having an attitude of humility, can provide people and communities resources to actively re-shape their interpersonal relationships in their communities to promote and sustain actions toward social change .Besides a need, research funding depends on an engaged funder. Government funding is likely if these social and economic, and racial pressures do not abate. Private funding from foundations and NGOs might also be aimed at these social tensions.New treatments inevitably provide, for some, an attraction of using a new technology to deliver the treatments. In addition, philosopher Peter Knowledge leads. Often, innovators are drawn to initial knowledge, usually combined with internal drives to be seen as opinion leaders. Early knowledge is crucial to innovators’ adoptions. Yet most of the innovations fail. Thus, when innovators lead, usually at least some empirical success must attend their choice.Persuasion Social confirmation. Once people have adopted a technology—whether innovator or laggard or anyone between—we all look for social confirmation that we have made the right choice. As cognitive psychologists tell us, we look to support our choices not disconfirm them .Because intervention research in temperance is in its infancy, the future looks rosy for PPI researchers. However, in this perspective paper, we suggest that we are in a second wave of PPI intervention research  that reqIn conclusion, to promote growth in temperance interventions, our lessons are few: First, new innovations must be developed—like creating temperance-related PPI apps, games, web-based brief interventions, engaging Internet-based interventions, and artificial intelligence interventions. Second, these innovations must be tested empirically for efficacy, effectiveness, dissemination, and scaling—each of which could necessitate modifications of content and delivery methods. Third, innovators must be attracted to the new methods, and the innovators must be likeable, have networks of loyal followers, and be persuasive enough to wean people from their existing commitments to temperance-focused evidence-based PPIs. Fourth, there must be initial positive data because the enthusiasm of innovators cannot carry a technology without some evidence of its efficacy. Fifth, leaders in the field must endorse the new intervention. Sixth, word-of-mouth communication of evidence that the new methods are successful in essential to build a following. Seventh, social proof and ultimately social confirmation are needed to sustain the use of the new innovations. Our fervent hope is that the present perspective article might inspire changes and provide ideas for new steps in building temperance by using PPIs.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.EW conceptualized the paper. EW and LEvZ drafted and finalized the first version of the manuscript. All authors contributed to the article and approved the submitted version.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Eurosurveillance issue, two articles by Colzani et al. [In this i et al.  and Ortui et al. , raise tColzani et al.  use the Among the more than 300 different HPVs infecting humans, types 16, 18, 31 and 45 are the most oncogenic viral agents for humans with over 90% of the HPV-associated cancer burden. The elimination of HPV-associated cancers will be accomplished by herd effect enforced eradication of these most oncogenic and prevalent HPV types. Because of both, the much lower R0, less than 4 for all other HPV types than HPV16, and the assortativeness of sexual risk-taking behaviour, it is the herd effect from moderate to high coverage gender-neutral vaccination that is the relevant factor providing protection even against HPV16 [Colzani et al.  meticuloMost importantly, it may not be possible to easily circumvent HPV vaccine shortage by changing the vaccination schedule or dosing. No HPV vaccine-induced protective antibody level has been defined and overall, the effect from one-dose vaccinations, aside from that with live vaccines, is unclear and assumes boostering which has its caveats (e.g. likely limited access to and availability of services) in the LMICs. Studies have shown that up to 10% of the 4-valent HPV vaccine recipients do not have neutralising or total HPV18 L1-VLP (virus-like particle) antibodies 7 to 12 years post vaccination , which e


The timely detection of precancerous lesions and early intervention can greatly reduce cervical cancer occurrence. The current study aimed to assess the diagnostic value and accuracy of different methods of cervical lesion screening. A total of 1622 females who visited the Outpatient Department of Xinjiang Uyghur Autonomous Region People’s Hospital between January and December 2018 were consecutively enrolled. All participants underwent separate high-risk human papilloma virus (HR-HPV) DNA detection, ThinPrep cytology testing (TCT) and colposcopic biopsy. Their medical records were retrospectively analyzed. While considering biopsy outcomes as the gold standard, the diagnostic values of TCT, HR-HPV testing, and TCT+HR-HPV testing for cervical cancer screening were compared. The sensitivity, specificity and Youden index of each method were calculated. Among the different methods, TCT+HR-HPV testing had the highest sensitivity (89.8%), followed by TCT (79.9%) and HR-HPV testing (49.2%). The combined method also had the highest Youden value, and its screening outcomes exhibited the highest consistency with those of biopsy. In addition, the combined method had the largest area under the receiver operating characteristic (ROC) curve, which was 0.673 , compared with any other screening method. Compared with TCT or HR-HPV testing alone, TCT+HR-HPV testing serves as a better screening method for cervical cancer and precancerous lesions. Cervical cancer is the fourth most common malignancy in terms of incidence and mortality in women worldwide. In 2018, a total of 570,000 new cases of cervical cancer were diagnosed worldwide, and the number of deaths due to cervical cancer reached 311,000; among cervical cancer-related deaths, approximately 85% occurred in developing and undeveloped countries, and the mortality was 18 times higher in low–middle-income countries than high-income countries [China is the largest developing country in the world, and cervical cancer is highly prevalent in China. Annually, approximately 130,000 new cases of cervical cancer are diagnosed in China, which accounts for 28.8% of all new cases worldwide; the annual number of cancer-related deaths in China is approximately 80,000, and the mortality ranks second only to that caused by breast cancer . In XinjThe progression of cervical cancer from cervical intraepithelial neoplasia (CIN) is a long, reversible process, and this period is referred to as the precancerous stage. If disease is detected at this stage  and timely intervention is then provided, the progression of CIN may be prevented, thereby reducing the possibility of cervical cancer occurrence .HPV infection is the primary cause of precancerous lesions and cervical cancer; high-risk HPV (HR-HPV) infection is the main risk factor for high-grade CIN and cervical cancer, which has been evidenced by research on both pathogenesis and vaccines , 10, 11.In this study, we compared the screening value and accuracy of ThinPrep cytology testing (TCT), HR-HPV DNA testing and TCT combined with HR-HPV testing (TCT+HR-HPV) in cervical cancer screening for women in Xinjiang. The results of this study may provide a fast, accurate cervical cancer screening method for the population of Xijiang, as well as for those in other regions with similar socioeconomic backgrounds.Between January and December 2018, a total of 2190 women who visited the outpatient office of the Department of Gynecology of Xinjiang Uyghur Autonomous Region People’s Hospital for cervicitis, intraepithelial neoplasia or cervical cancer or only for cervical cancer screening were considered for inclusion in this study. They were informed of the research goals and voluntarily completed a series of separate examinations, including HR-HPV testing, TCT and pathological biopsy. Their ages ranged from 15 to 65 years old. All participants met the following inclusion criteria: 1) aged between 18 years and 65 years; and 2) a history of sexual intercourse (based on inquiring whether the patient had such a history as well as observing the hymen condition at the time of sample collection). The exclusion criteria were as follows: 1) pregnancy or pregnancy termination within 3 months prior to this study; 2) refusal to receive colposcopic biopsy or dysplasia of the cervix according to colposcopic biopsy at the time of the current visit; 3) no history of sexual intercourse; 4) mental disorders or other severe diseases; 4) uterine, cervical or vaginal hemorrhage at the time of diagnosis; 5) acute lower genital tract, vulvar, vaginal or cervical infection; and 6) other concurrent sexually transmitted diseases. Finally, 1622 females qualified for this study. Their medical records included general and clinical data and were retrospectively analyzed between December 1, 2019, and December 31, 2019. A flow chart describing recruitment is shown in The procedures of this study were approved by the Ethics Committee of Xinjiang Uyghur Municipal People’s Hospital (KY20180118164). Written informed consent for research purposes was obtained from each participant.Exfoliated cells at the external aperture of the cervix and the cervical canal were collected with a sampling brush and then stored in a bottle with preservation solution. A thin-layer cell smear was made with a ThinPrep 2000 system for Pap examinations. Diagnoses were made according to the grading criteria of the Bethesda System: 1) no intraepithelial or malignant lesions (NILM); 2) squamous epithelial cell abnormality: a, atypical squamous cells (ASCs), including ASC-US and atypical squamous cells that cannot exclude high-grade squamous intraepithelial lesion (ASC-H); b, low-grade squamous intraepithelial lesion (LSIL); c, high-grade squamous intraepithelial lesion (HSIL); and d, squamous cell carcinoma (SCC); 3) glandular epithelial cell abnormality: a, atypical glandular cells (AGCs), including AGC-not otherwise specified (AGC-NOS) and AGC-suspicious for neoplasia (AGC-N); b, cervical adenocarcinoma in situ of the cervical canal (AIS); and c, adenocarcinoma; and 4) other malignant tumors.5’-CGTCCMARRGGAWACTGATC-3’ (MY09) and 5’-GCMCAGGGWCATAAYAATGG-3’ (MY11), and the primers for β-globin amplification were 5’-ACA CAA CTG TGT TCA CTA GC-3’ (F) and 5’-CAA CTT CAT CCA CGT TCA CC-3’ (R). Viral genes and types were determined using reverse dot-blot (RDB) hybridization . According to the criteria indicated in the kit, HR-HPV >1.0 pg/ml was considered positive.Viral DNA was amplified using real-time fluorescent quantitative polymerase chain reaction (RT-PCR), and the procedures were performed with the HPV genotyping kit with specificity of 98%, accuracy of 98% and repeatability of 98% (Yaneng Biosciences (Shenzhen) Co., Ltd., China). The universal primers for fragment amplification of HPV L1 genes were Colposcopy was performed after coating with 3% acetic acid. Iodine staining was performed. For each participant, four tissue samples were taken from the iodine-uncolored regions for histopathological examination . For those with suspected lesions, based on the outcomes of acetic acid coating and iodine staining, extra tissue was taken from the abnormal site. The pathological diagnosis criteria for CIN and carcinoma in situ were as follows: 1) CIN-I, mild dysplasia; 2) CIN-II, moderate dysplasia; 3) CIN-III, severe dysplasia, including carcinoma in situ; 4) carcinoma in situ and early infiltration; and 5) infiltrating carcinoma, including squamous carcinoma and adenocarcinoma.The judgment criteria for the results of different examination methods were as follows:TCT: all results were positive except for NILM results;TCT+HR-HPV: if the result of either method was positive, the final result was considered positive; a final result was considered negative only when the results of both methods were negative.If a patient was diagnosed with ASC-US or LSIL according to TCT and with LSIL or other benign lesions according to pathological biopsy, the TCT outcome was considered “consistent”. If the TCT outcome was the same but the patient was diagnosed as HSIL or a condition more severe than HSIL, the TCT outcome was considered an “insufficient diagnosis”. If the TCT outcome for a patient was ASC-H, HSIL or SCC and that of pathological biopsy was HSIL or SCC, the TCT outcome was considered “consistent”. However, if the TCT outcome remained the same but the pathological biopsy outcome was LSIL or a condition less severe than LSIL, the TCT outcome was considered an “overdiagnosis”. Both “insufficient diagnosis” and “overdiagnosis” were considered an “inconsistent diagnosis”.Data were processed with SPSS 21.0. While considering the outcomes of colposcopic biopsy to be the gold standard, the diagnostic values of TCT, HR-HPV DNA testing and TCT+HR-HPV testing were compared with the chi-square test. The sensitivity, specificity, Youden index and consistency of each approach were determined. A difference of P<0.05 was considered statistically significant.A total of 1622 females received simultaneous TCT, HR-HPV testing and colposcopic biopsy. Their baseline information is summarized in The detection rates of TCT for CIN-I, CIN-II, CIN-III, and cervical cancer were 64.7% (154/238), 88.3% (309/350), 81.3% (122/150), and 100.00% (24/24), respectively ; S1 FileThe detection rates of HPV testing for CIN-I, CIN-II, CIN-III, and cervical cancer were 38.2% (91/238), 50.3% (176/350), 59.3% (89/150), and 79.2% (19/24), respectively ; S1 FileThe detection rates of TCT+HPV testing for CIN-I, CIN-II, CIN-III, and cervical cancer were 86.5% (206/238), 94.0% (329/350), 83.3% (125/150), and 100% (24/24), respectively ; S1 FileThe diagnostic values of the three screening strategies were then compared. Among them, HR-HPV testing had the highest specificity (64.5%) but the lowest sensitivity (49.2%). In contrast, TCT+HR-HPV testing achieved a sensitivity of 89.8%. Furthermore, the Youden index and consistency of TCT+HR-HPV testing were both higher than those of any other strategy . In addiAlthough TCT and HPV testing have been widely applied in cervical cancer screening in China, comparisons of the diagnostic value of different screening methods based on a large sample size have rarely been reported. In this study, we compared the diagnostic values of TCT, HR-HPV testing and TCT+HR-HPV testing in a sample size of 1622 patients.According to this study, the Youden index and consistency level of TCT+HR-HPV testing were higher than those of any other method. In the meantime, the AUC for TCT+HR-HPV testing was also the largest among the three assessed methods. These findings indicate that TCT+HR-HPV testing has the highest outcome accuracy, followed by TCT. In terms of sensitivity, TCT+HR-HPV testing had the most powerful ability to accurately identify cervical cancer, whereas HR-HPV testing had the weakest ability. In addition, our study showed that HR-HPV testing had the highest specificity, whereas TCT had the lowest specificity, which indicates that HR-HPV testing has the most powerful capacity for accurate noncervical cancer screening.Precancerous lesion detection acts as a critical early warning for the risk of cervical cell cancerization and for the timely prevention and treatment of cervical cancer at an early stage. This study showed that although TCT had a low CIN-I detection rate, its detection rates for CIN-II and above were high (both exceeding 80%), particularly for cervical cancer, which reached 100%. HR-HPV testing had relatively low detection rates for CIN-I through CIN-III, and its detection rate for cervical cancer was lower than 80%. TCT+HR-HPV testing had high detection rates for CIN-I through CIN-III, and its detection rate for cervical cancer reached 100%. These findings indicate that TCT+HR-HPV testing is highly valuable for precancerous lesion screening. Long et al.  comparedCervical epithelial cells undergo a series of pathological changes before cervical cancer occurs; effective screening during this period can greatly reduce the incidence rate of the malignancy . The WHOIn this study, the HR-HPV positivity rate exceeded 40%, which was noticeably higher than that reported by Wang et al. . In WangThis study had some limitations. First, this was a single-center study. Therefore, to verify the results of this study, multicenter studies need to be conducted in the future. Second, this study focused on the efficacy of different screening methods rather than investigating the prevalence rate of cervical cancer in the involved region. Furthermore, the included individuals were all from our hospital; that is, the samples were not based on the whole population in the involved region. Therefore, the obtained prevalence rates of HPV positivity and cervical cancer in this study may not represent those of the involved region. Third, despite the positive results obtained in this study, laboratory tests only serve as adjuvant measures for clinical diagnosis, and cervical diseases can only be confirmed according to clinical symptoms and pathological examination.In conclusion, TCT+HR-HPV testing improves the accuracy of cervical lesion screening in clinical practice. Therefore, it can be popularized and integrated as management guidelines in Xinjiang, China, where the incidence of cervical cancer is high. Furthermore, it can be used as an important screening approach for cervical cancer and precancerous lesions.S1 File(XLSX)Click here for additional data file.


Human papillomavirus (HPV) infection identified as a definitive human carcinogen is increasingly being recognized for its role in carcinogenesis of human cancers. Up to 38%–80% of head and neck squamous cell carcinoma (HNSCC) in oropharyngeal location (OPSCC) and nearly all cervical cancers contain the HPV genome which is implicated in causing cancer through its oncoproteins E6 and E7. Given by the biologically distinct HPV-related OPSCC and a more favorable prognosis compared to HPV-negative tumors, clinical trials on de-escalation treatment strategies for these patients have been studied. It is therefore raised the questions for the patient stratification if treatment de-escalation is feasible. Moreover, understanding the crosstalk of HPV-mediated malignancy and immunity with clinical insights from the proportional response rate to immune checkpoint blockade treatments in patients with HNSCC is of importance to substantially improve the treatment efficacy. This review discusses the biology of HPV-related HNSCC as well as successful clinically findings with promising candidates in the pipeline for future directions. With the advent of various sequencing technologies, further biomolecules associated with HPV-related HNSCC progression are currently being identified to be used as potential biomarkers or targets for clinical decisions throughout the continuum of cancer care. The incidence of HPV-related head and neck squamous cell carcinoma (HNSCC), especially oropharyngeal squamous cell carcinoma (OPSCC), has increased in the United States and Europe  degradation, consequently leading to a deregulation of the cell cycle and an inhibition of apoptosis  has been reported in cervical cancer and HNSCC , there are more than 225 distinct genotypes of HPVs, which are classified according to the DNA sequence of the L1 gene. To be recognized as a new genotype, the L1 region sequence has to be more than 10% different from its closest member. Despite the sequence variation, the HPV genome structure is similar for all genotypes and is organized into three regions  lead to the activation of the PI3K-AKT-mTOR1 signaling pathway necessary for the viral life cycle , Inc-FANCI-2, mediated by E7 and E6, which is independent of p53/E6AP and pRb/E2F in cervical carcinogenesis  between HPV– and HPV+ HNSCCs by single-cell transcriptional profiling   staining, quantitative polymerase chain reaction (qPCR)-based HR-HPV DNA or RNA testing, and HPV DNA or RNA in situ hybridization (ISH) have been established. In 2018, the College of American Pathologists (CAP) and American Society of Clinical Oncology (ASCO) released guidelines that HR-HPV testing should be performed for OPSCC and cervical nodal metastases of unknown primary tumors  of HPV-related HNSCC has been identified as contributing to the pathophysiology of the disease. This variability contributes to the range of treatment responses observed for both established clinical practice and clinical trials. The clinical translation of these findings may help for dynamic risk restratification of HPV-related HNSCC patients with new molecular biomarkers. The management of HPV-related HNSCC is changing, and substantial research focusing on discovery approaches to cancer diagnostics and prognostic evaluations is required (Bigelow


Aberrant microbiota composition has been linked to disease development at numerous anatomical sites. Microbiota changes in reaction to viral infections, such as human papillomavirus (HPV), have been investigated almost exclusively in the female reproductive tract. However, HPV infection may also affect male health by reducing semen quality and fertility. The aim of this study was to investigate whether present HPV DNA is associated with detectable changes in semen bacterial microbiota composition and diversity.This study relied on stored semen samples from 31 fertile healthy men who participated in the Finnish family HPV Study during the years 1998–2001. DNA was extracted from semen with PCR template preparation kit. HPV was genotyped using Luminex-based Multimetrix® assay. Microbiota was analyzed from the V3-V4 region of 16S rDNA gene following sequencing on an Illumina MiSeq platform. All statistical analyses were performed with Calypso software version 8.84.Moraxellaceae (p = 0.028), Streptococcus (p = 0.0058) and Peptostreptococcus (p = 0.012) compared to HPV-negative semen samples.HPV DNA was detected in 19.4% (6/31) of the semen samples. HPV status in the semen did not impact the α-diversity estimations, as measured by Chao1 and Shannon indices, nor ß-diversity. Nevertheless, HPV-positive semen samples exhibited differences in the taxonomic composition of the bacterial microbiota including higher abundances of HPV infection is associated with altered bacterial microbiota composition in semen, and this might have in impact to male health in general. As of present, it is unclear whether these changes result from HPV infection or whether altered bacterial microbiota increases susceptibility to HPV infection. More research is needed on viral-bacterial interactions in the male reproductive system.The online version contains supplementary material available at 10.1186/s12879-021-06029-3. Human papillomavirus (HPV) is a known oncovirus and it has potential to cause carcinoma in the genital, anal and oropharyngeal regions . The natSeveral studies have reported the presence of HPV DNA in 2–31% of semen samples from healthy men –7. LuttmLactobacillus, Pseudomonas, Prevotella, Gardenella, Corynebacterium, Staphylococcus and Streptococcus [Although limited, published literature in the last decade have given us a glimpse into the composition of semen microbiota. The semen microbiota is mainly composed of the genera tococcus –17. Semetococcus , 14, 16.tococcus , 16, 18.tococcus , 14. HowSince it is known that seminal microbiota can influence fertility and even offspring health, it is important to understand aspects that might affect semen microbiota composition . We inveThe subjects and samples of the present study were from a sub-set of subjects from the Finnish Family HPV Study collected during the years 1998–2001 . The ori, Table Semen samples for microbiota analyses were collected based on the placenta HPV status as previously described in detail . The ori1Semen samples were collected during the female spouse’s third trimester. The detailed sample collection is described earlier . Semen sDetailed description about the semen HPV DNA detection and genotyping is described earlier . HPV DNAhttp://cgenome.net/calypso/) [p-values ≤0.05 statistically significant.Sample analyzing methods have previously been described in detail , 3, 22. alypso/)  was usedHPV DNA was detected in 6 of the 31 semen samples (19.4%). Infection with a single HPV genotype was found in five of the samples , as seen in Table None of the HPV DNA positive semen samples associated with female spouse’s HPV status in oral, cervix or from the placenta as we did not observe the same HPV genotype in partners simultaneously.Comamonadaceae (relative abundance 4.13%), Bifidobacteriaceae (2.44%), Tissirellaceae (2.39%) and Corynebacteriaceae (2.11%). In addition, Delftia (3.76%), Unclassified Comamonadeceae (1.56%), Propionibacterium (1.55%) and Streptococcus (1.52%) genera were detected. The most prevalent bacteria detected from HPV-negative and HPV-positive semen samples are presented in Supplementary Table The predominant bacterial families detected in the semen samples consisted of eriaceae .44%, Tisp = 0.820, Fig. p = 0.058, Fig. p = 0.494) or redundancy analysis  as presented in the Supplementary Data HPV status in semen did not impact the alpha diversity estimations, as measured by two alpha diversity metrics: Chao1 , Peptostreptococcaceae , Veillonellaceae  and Moraxellaceae  were more abundant in the HPV-positive samples than in HPV-negative samples , Serratia , Dialister  and Peptostreptococcus  compared to HPV-negative semen samples , Anaerococcus mediterraneensis  and Streptococcus anginosus  were observed in the HPV-positive semen group relative to the HPV-negative group.Since there was no major impact to semen bacterial microbiota composition in general according to HPV status, we sought to examine if there were any differentially abundant taxa in HPV-negative and HPV-positive semen samples. Indeed, semen HPV status was associated with differences in the relative abundance of specific bacterial taxa. At phylum level, Fusobacteria were more abundant in HPV-negative (relative abundance: 0.450%) compared to HPV-positive 0.032%) samples, % samplesComamonadaceae, Bifidobacteriaceae, Tissirellaceae, Corynebacteriaceae, Delftia, unclassified Comamonadeceae, Propionibacterium and Streptococcus members. The overall seminal bacterial composition has been seen to vary between different studies [Lactobacillus, Corynebacterium, Streptococcus, Pseudomonas, Prevotella and Gardenella mainly constitute the healthy/normal semen bacterial microbiota [. The bacterial composition in our semen samples were detected to be somewhat different compared to previous studies, but overall, the genera Corynebacterium and Streptococcus were consistent. The small sample size may have an effect on our results.In the present study, we detected that the presence of HPV in semen is associated with some detectable changes in the semen bacterial microbiota composition in healthy individuals. The bacterial microbiota of the semen samples consisted mainly of  studies , 14. Precrobiota , 14, 17.The semen microbiota can be influenced by the bacteria colonized the penis, urethra and from the urine , 26–30. In the HPV-negative samples we detected higher abundance of Fusobacteria when compared to HPV-positive semen. Fusobacteria is a known oral pathogen  which haGiven that semen HPV and microbiota has been implicated in seminal health and disease/disorders, including infertility , 19, 33 Streptococcaceae, Peptostreptococcaceae, Veillonellaceae, in addition with Streptococcus, Serratia, Dialister and Peptostreptococcus  [In our study, we detected HPV-positive semen samples to be more abundant with nt study . Furtherlonella) .Lactobacillus, Streptococcus, Veillonella and Peptostreptococcus with undefined male infertility [, the abundances of Lactobacillus and Streptococcus have been identified to be significantly higher in semen samples from healthy men [Streptococcus and Peptostreptococcus genera as compared to HPV DNA negative semen. Among the bacterial microbiota found in semen, Ureaplasma has been described as a known pathogen connected strongly with male infertility [Anaerococcus in semen have been associated in inferior semen quality in previous studies [Previous studies have connected ertility , 39. Whilthy men , 14. In ertility . Both HP studies , 14, 36. studies . NeverthPreviously, it has been shown that present viral infection with HIV can shape the bacterial microbiota composition and that the altered bacterial composition might enable viral transmission in addition with changes in the pro-inflammatory cytokines . It coulThis study has some limitations. Our study design restricts us from making definitive conclusions about the viral-bacterial associations. Furthermore, the relatively low number of samples in general and the low number of HPV-positive semen samples may have an impact on the results, as we did not find significant impact of seminal HPV on the diversity of semen microbiota. HPV DNA may also originate from the proximal excurrent ducts  and not from HPV-infected seminal cells , 41. TheIn this study, we showed that men with seminal HPV DNA had higher relative abundance of specific seminal bacteria compared to men without seminal HPV DNA, but the causal relationship remains to be determined in future studies. Furthermore, data is largely lacking on whether the interplay between seminal HPV and semen microbiota has any clinical significance in male reproductive health.Improvement of semen quality and securing healthy semen microbiota may be among the beneficial effects of HPV vaccination. Recently, HPV vaccination has been shown to improve the clearing of semen HPV infections and the reproductive outcomes in infertile patients with semen HPV infection .Additional file 1:Table 1. The list of the samples.Additional file 2:Supplementary Table 2. A) The most prevalent bacteria detected in HPV-negative semen samples. B) The most prevalent bacteria detected in HPV-positive semen samples.Additional file 3:Supplementary Data 1. The beta-diversity estimations .


In this study, the cause of failure of a low-carbon steel pipe meeting standard KS D 3562 (ASTM A135), in a district heating system was investigated. After 6 years of operation, the pipe failed prematurely due to pitting corrosion, which occurred both inside and outside of the pipe. Pitting corrosion occurred more prominently outside the pipe than inside, where water quality is controlled. The analysis indicated that the pipe failure occurred due to aluminum inclusions and the presence of a pearlite inhomogeneous phase fraction. Crevice corrosion occurred in the vicinity around the aluminum inclusions, causing localized corrosion. In the large pearlite fraction region, cementite in the pearlite acted as a cathode to promote dissolution of surrounding ferrite. Therefore, in the groundwater environment outside of the pipe, localized corrosion occurred due to crevice corrosion by aluminum inclusions, and localized corrosion was accelerated by the large fraction of pearlite around the aluminum inclusions, leading to pipe failure. District heating (DH) systems produce heat that is used to provide steam and hot water to the residents of large cities ,2. TheseLow-carbon steel has been widely used as a pipe material in various industrial plants such as those that manage oil, gas, and water, as well as in DH systems ,5,6,7,8.In most studies, the failure of low-carbon steel pipe occurs in the form of stress corrosion cracking (SCC) or corrosion fatigue cracking (CFC) when stress is applied. However, the failure shown in this study was due to pitting corrosion. Low-carbon steel is not passivated as stainless steel is, and corrosion generally occurs uniformly over the pipe. Pitting corrosion is rarely observed on the low-carbon steel pipe . TherefoThis study analyzes the failure of a low-carbon steel pipe due to corrosion, which has not been reported in the actual use of low-carbon steel pipe. Compliance with material specification was evaluated using inductively coupled plasma atomic emission spectroscopy (ICP-AES) component analysis. The cause of the failure was then analyzed through visual inspection, optical microscopy (OM), metallographic examination, scanning electron microscopy (SEM) with energy dispersive spectroscopy (EDS), electron probe microanalyzer (EPMA) and atomic force microscopy (AFM). Furthermore, the electrochemical properties of the failed low-carbon steel were evaluated using the potentiodynamic polarization and galvanostatic polarization tests.Before conducting the failure analysis of the low-carbon steel pipe, the specifications and environment were investigated. According to the user’s description, the failed low-carbon steel pipe transported hot water through underground pipeline in DH systems. The low-carbon steel pipe satisfied standard KS D 3562  ,13,14. Trms at 17 kHz. Measurements were performed at 10 µm × 10 µm and 2 µm × 2 µm, respectively. The fracture properties were analyzed using SEM/EDS  and EPMA . Prior to SEM/EDS and EPMA analyses, specimens were pickled, polished with 1000-grit size silicon carbide paper, and then rinsed with deionized water and cleaned with ethanol.In this study, it was investigated whether the material used for the pipe satisfies the KS D 3562 standard through chemical composition analysis. The shape of the failed pipe was visually investigated via OM. The metallographic examination was performed to confirm the microstructure uniformity of the pipe metal. To investigate the microstructure of the pipe metal, the specimen was polished using a 1-µm diamond suspension, and thereafter etched by a 2% Nital etching solution for 20 s . The vol2 using a sealant. A groundwater solution was used for the electrochemical tests (corr). The potentiodynamic polarization tests were conducted using a potential sweep of 0.01 mV/s from −0.25 V vs. Ecorr to 1.6 V vs. Ecorr. The galvanostatic polarization tests were performed at 5 mA/cm2 to accelerate corrosion. The total amount of coulombic charge was 143.86 mAh, which is equivalent to 6 months of uniform corrosion in specimen B. To evaluate the same amount of corrosion, the same coulombic charge was applied to both specimens.Potentiodynamic polarization and galvanostatic polarization tests were performed using a VSP 300 . To conduct these electrochemical tests, a three-electrode system comprising low-carbon steel pipe specimen taken from the failed pipe as the working electrode (WE), two pure graphite rods as the counter electrodes (CE), and a saturated calomel electrode (SCE) with a Luggin capillary as the reference electrode (RE) was used. To confirm the cause of the pitting corrosion, specimens were prepared for the pitting corrosion part (specimen A) and the uniform corrosion part (specimen B) of the failed pipe. For electrochemical tests, all specimens were polished with a 1000-grit silicon carbide paper, rinsed with ethanol, and dried with nitrogen gas. The area of all specimens was controlled to a size of 1 cmal tests  because Based on visual inspection of the failed pipe, leakage occurred in the form of pitting, and the severe pitting corrosion occurred near the part of the pipe where the leaking occurred . Figure Moreover, the depth of the pitting corrosion on the exterior of the failed pipe was significantly deeper than the interior. It is considered to have been due to the difference between the environment of the inside and outside of the pipe . In addiTherefore, the inhomogeneity of the microstructure forces a difference in corrosion rate. A pipe section exhibiting pitting corrosion with a lot of pearlite experiences severe corrosion, whereas a uniform corrosion section of the failed pipe with a low pearlite phase incurs only minor thickness reduction.To identify the cause of severe pitting corrosion, microscopic analyses were performed on the pitting corrosion near the leakage area using SEM/EDS and EPMA . Figure Al inclusions were considered to have been the result of Al use as a deoxidizer in the steelmaking process . Al is kcorr than specimen B, and had a relatively large potential fluctuation of approximately 50 mV. Specimen A has a higher Ecorr due to a larger fraction of pearlite, which has a relatively noble potential compared to ferrite. Pearlite has a higher Ecorr than ferrite due to an increase in cathodic sites that cause oxygen reduction (O2 + 2H2O + 4e = 4OH−) [ = 4OH−) ,32. In a = 4OH−) ,34.− ion, are concentrated around the Al inclusion. This further accelerates crevice corrosion in Al inclusions. If there is no crevice around the Al inclusions, crevice corrosion does not occur. In addition, when pitting formed as crevice corrosion and galvanic corrosion progressed around the Al inclusions and the pearlite, the surface area became wider than the initial area due to morphological changes [ion rate . Larger ion rate . In addi changes ,35. Equa changes .(2)ia=iIn Equation (2), 2 for the generated current (In Equation (3), 2), 2. The galvanostatic polarization test was performed to accelerate corrosion. The acceleration time for the galvanostatic polarization test was calculated using the Faraday’s law as shown below :(4)ireal− ions are concentrated in the microcrevice to maintain charge neutrality [− ions further accelerates crevice corrosion and corrosion products accumulate on this part of the pipe. Additionally, oxygen-concentration cells form, which accelerate localized corrosion, and eventually the Al inclusions fall off [utrality . The confall off ,29.− ions into Al vicinity and the accumulation of corrosion products are accelerated. This promotes the formation of an oxygen-concentration cell in the vicinity of the Al inclusion.The large fraction of pearlite has a higher corrosion rate due to microgalvanic corrosion between the surrounding pro-eutectoid ferrite and pearlite, and between cementite and ferrite in pearlite ,24. Due However, the presence of a large fraction of pearlite alone cannot cause localized corrosion such as pitting. The cementite inside the pearlite locally accelerates the surrounding ferrite corrosion by microgalvanic corrosion, but over time, the rust is covered by the dissolution of ferrite, and the corrosion proceeds in the form of a uniform corrosion . In otheIn this study, the failure analysis of a low-carbon steel pipe used in DH system was investigated using visual inspection, ICP-AES, OM, AFM, SEM/EDS, EPMA, and electrochemical tests. According to the results of the failure analysis, the following conclusions were drawn.Leakage occurred in the form of pitting corrosion, which was observed both inside and outside of the failed pipe. In particular, severe pitting corrosion occurred on the outside of the pipe, exposed to the soil environment. Al inclusions and a larger phase fraction of pearlite were observed near the leaking section. Crevice corrosion occurred in the microcrevice around the Al inclusions, and the large phase of pearlite around Al inclusions accelerated the localized corrosion in the microcrevice. Localized corrosion was accelerated near the Al inclusions and the large fraction of pearlite in the groundwater environment outside of the pipe, resulting in the pipe’s failure.The corrosion rate of the specimen taken where the pitting corrosion was present in the failed pipe was approximately double that of the specimen taken from the uniform corrosion part of the failed pipe. Furthermore, the corrosion type was similar to that observed in the actual failed pipe. This confirms the pipe failure had been caused by Al inclusions and the inhomogeneity of the pearlite.It is recommended that the uniform distribution of fine-sized pearlite and Al inclusions be produced through proper liquid steel homogenization and heat treatment during the steelmaking process.It is recommended that a standard for the chemical composition of Al be established within the existing KS D 3562 standard.


The blue phases are observed in highly chiral liquid crystalline compositions that nascently organize into a three-dimensional, crystalline nanostructure. The periodicity of the unit cell lattice spacing is on the order of the wavelength of visible light and accordingly, the blue phases exhibit a selective reflection as a photonic crystal. Here, we detail the synthesis of liquid crystalline elastomers that retain blue phase I, blue phase II, and blue phase III. The mechanical properties and optical reconfiguration via deformation of retained blue phases are contrasted to the cholesteric phase in fully solid elastomers with glass transition temperatures below room temperature. Mechanical deformation and chemical swelling of the lightly crosslinked polymer networks induces lattice asymmetry in the blue phase evident in the tuning of the selective reflection. The lattice periodicity of the blue phase elastomer is minimally affected by temperature. The oblique lattice planes of the blue phase tilt and red-shift in response to mechanical deformation. The retention of the blue phases in fully solid, elastomeric films could enable functional implementations in photonics, sensing, and energy applications. Permanently retaining liquid crystalline blue phases in a solid matrix warrants many prospective applications in photonics. Schlafmann and White are able to make a blue phase rubber that retains a crystalline structure upon deformation and responds with angle-dependent color changes. Recently, they have drawn considerable research interest for their utility as machine-like mechanical actuators2 as well as the spatial programmability of their material properties3. Analogous to small-molecule liquid crystalline systems, which have been prevalent in display applications for some time, LCEs also are relevant to a variety of optical and photonic applications including solid-state optical sensing4, tunable diffraction5, and adaptive lasing6.Liquid crystalline elastomers (LCEs) are compelling responsive materials7. In certain functional implementations, the realization of dynamic optical response in fully solid films is particularly desirable. Further, as evident in biological systems, dynamic reconfiguration of solid skins naturally induces multifunctionality even under ambient conditions8. Ranging from regulated structural color and diffuse reflectance in cephalopods10 to glare-reducing corneal nipple arrays on nocturnal insect eyes12 and photonic crystalline structures responsible for coloring in butterfly wing scales14, the natural environment provides countless examples of dynamic light control. Many synthetic material systems have incorporated bio-inspired mechanisms to imitate their photonic properties for technological applications15.Here, we detail an approach to prepare fully solid photonic materials that are robust, yet elastic and capable of dynamic reconfiguration. The fluidic nature of small-molecule liquid crystalline systems enables the response time and performance of liquid crystal displays16. These unique periodic nanostructures exhibit photonic band gaps in multiple propagation directions. Three-dimensional photonic crystals are nanoscale emulators of crystal structures typically observed in atomic lattices. Nanostructures such as those arising from inverse opals or lithographic deposition exhibit three-dimensional photonic crystallinity, however, the intensive fabrication of such materials is complex and costly17. The optics and photonics community has increasingly looked towards self-assembly and liquid crystals as light-manipulating media. Liquid crystalline materials can be formulated to adopt a catalog of anisotropic phases. Chiral liquid crystalline phases in particular exhibit photonic band gaps due to their periodic anisotropy18.Multidimensional photonic crystals such as those observed in the structural coloration in certain animals are of particular interest for optical applications such as nonlinear optics and waveguides19. These cylinders stack upon one another to generate cubic lattices stabilized by periodic disclinations. Blue phase I (BPI) organizes as a body-centered cubic (bcc) lattice, blue phase II (BPII) achieves a simple cubic (sc) lattice, and blue phase III  is largely amorphous. In this way, BPI and BPII are three-dimensional photonic crystals that are selectively reflective to visible light19.The liquid crystalline blue phases are a subset of liquid crystalline phases in which calamitic mesogens align in a double-twist morphology. The blue phases are frustrated phases and the nanostructure of these materials are a combination of defect packing and double-twist molecular arrangement of cylinders on the order of 100 nm in diameter27. The primary motivation for these studies is the potential for fast electro-optical switching properties in devices that do not need alignment layers or precise thickness control28. They have also been retained in free-standing glassy polymers upon cross-linking within the thermotropic phase window34. These polymers exhibit largely static photonic properties due to their high cross-link density, however recently the selective reflections of these materials have been sensitized to humidity and pH through supramolecular bonds within the network33 and to small amounts of mechanical deformation34. Polymer-stabilized blue-phase gels (32 wt.% polymerizable media) exhibit electro-optical response, along with limited photonic sensitivity to mechanical deformation35. However, the liquid crystalline blue phases have not been retained in truly elastomeric polymer networks with a glass transition below room temperature (Tg <20 °C).The cubic blue phases have been extensively studied in non-cross-linked low molar mass systems36. Here, enabled by similar reaction chemistry, we prepare liquid crystalline elastomers that retain the blue phases upon photopolymerization. We characterize changes in the lattice spacing to mechanical deformation, heat, and chemical exposure.We have reported a straightforward and scalable approach to prepare well-aligned cholesteric liquid crystal elastomers (CLCEs) in the planar orientation composed of main-chain chiral mesogens39 and/or anisotropic deswelling40. However, the intricate, three-dimensional nanostructure of the blue phases has not been retained in LCEs by these methods. A recent report36 details the development of a materials chemistry to prepare LCEs that originate from the desired chiral phase that are conducive to surface-enforced alignment and prepared from a one-step reaction. This chemistry was utilized to prepare and retain the cholesteric phase in LCEs of high optical quality (low haze) and considerable thermochromism.Historically, liquid crystalline elastomers (LCEs) have been prepared by two-stage polymerization reactions in which alignment is enforced in the second stage by mechanical force. The preparation of hierarchical liquid crystalline phases, such as the cholesteric or blue phases, are not readily amenable to these processes. The helicoidal nanostructure of the cholesteric phase has been retained in LCEs by mechanical alignment coupled with anisotropic deswelling through either centrifugation41. Polymerization is photoinitiated with Omnirad 819. A representative DSC trace of the LCE is illustrated in Fig. Tg) of ~15 °C. The acrylate-thiol chain transfer reactions suppress the cross-link density of the acrylate homopolymerization by producing a highly branched polymer network architecture illustrated in the inset to Fig. 1b41.The blue phases are observed in formulations that have a high concentration of chiral species. The liquid crystalline molecules organize in cubic, nanostructured phases. The periodicity of the blue phases is defined by crystalline packing within unit cells. The blue phases are typically observed in very narrow temperature ranges, which can complicate the retention of these phases in polymer networks. Here, we report the synthesis of LCEs that retain blue phase I (BPI), blue phase II (BPII), and blue phase III (BPIII). The composition is based on a mixture of the liquid crystalline diacrylate C6M and chiral liquid crystalline diacrylate SLO4151 Fig. . The non24. As expected, the cholesteric, BPIII, and polydomain nematic phases do not exhibit a diffraction pattern . Thus, we conclude the phase of the LCE is the primary differentiator in stress–strain behavior reported here.Informed and enabled by the retention of the blue phases in fully solid and elastomeric LCE, we focus the remainder of this examination on LCE retaining the BPI and BPII phases. The selective reflection of unperturbed BPI and BPII LCE, visualized in the photographs in Fig. 42 and observed43 in CLCE, owing to asymmetry in the period distribution of refractive index associated with thickness-dependent (orientation-dependent) reorientation to the strain axis. The biaxial strain of BPLCE results in an alternating pattern of yellow and blue regions, indicating a complex rearrangement of the mesogens coupled with the photonic properties of the blue phase. The physical reason for this distinctive texture is subject to further study.The mechanooptical response of the deformation of BPI and BPII was examined by UV/Vis spectroscopy during an applied load. The selective reflection  is incident upon the blue-phase LCE through a high numerical aperture objective Fig. . The reaEvident in Fig. The BPII LCE (sc) maintains [100] lattice alignment in the viewing direction with Kossel lines corresponding to the [010] and [001] planes observed at the expected angle of 90°. Similarly, the Kossel diagrams of BPII LCE during uniaxial deformation confirm that the lattice spacing becomes asymmetric to deformation. The simple cubic [100] periodicity decreases with strain. In an analogous fashion to the deformation-induced lattice tilting of BPII LCE, the [010] or [001]  tilts towards the film surface and maintains a periodicity much larger than that of the normal incidence [100] in the deformed state.θ) as the viewing angle increases. To further explore the spectral effect of mechanical deformation of the lattice, Fig. θ) dependence is equivalent. Again, the extraction of the nonreactive chiral dopant after the preparation of these films may impart some residual stress that could be the source of the lattice tilt that may be the origin of the small reflection apparent at 460 nm. The photographs of the unstrained BPII LCE in Fig. Again, the liquid crystalline blue phases are cubic structures; their selective reflection originates from the periodicity of that cubic lattice. In that sense, each set of lattice planes act as a Bragg reflector which blue-shifts ∝ cos, an alignment cell was prepared and selectively irradiated with a 405 nm laser  resulted in a comparatively larger uptake than the polar solvent acetone, which is qualitatively evident in the magnitude of the shift in reflection color. Exposure to dichloromethane resulted in even greater shifts into the near-infrared spectrum  deformation or asymmetric  deformation of the lattice of the blue phases, in all cases the distortion is reversible. The distinctive hierarchical nanostructure retained in the blue-phase LCE are unexplored but compelling for functional use as a lightweight, fully solid, and deformable optical materials of relevance in nonlinear optics, lasing, spectral imaging, and sensing.The achiral diacrylate C6M (Wilshire Technologies), the right-handed chiral diacrylate monomer SLO4151 (Alpha Micron), and the right-handed chiral dopant R811  were used as received. Blue-phase LCE mixtures were prepared by adding 51 wt% C6M, 19 wt% SLO4151, 15 wt% R811, 14 wt% BDMT , and 0.5 wt% Omnirad 819 (IGM Resins).2 for 10 min. Spatial variation in alignment utilized a photomask.Alignment cells were made by spin coating two Corning EXG glass slides with 22 μL of photoalignment dye PAAD-22 (Beam Co.) diluted (2 to 1 by volume) with DMF. The coated glass slides were dried at 100 °C for 30 min to drive off any residual solvent. Once dried, the coated sides of two glass slides adhered together with an epoxy adhesive mixed with 20-μm glass spacers. Photoalignment was achieved by irradiating the alignment cell with a linearly polarized 445 nm laser at an intensity of 10 mW/cm2. The polymer networks were extracted from the alignment cell and subsequently washed with acetone to remove any unreacted components.To prepare the liquid crystal elastomers retaining the various phases detailed hereto, the monomer mixture was melted to an isotropic phase at 90 °C and filled via capillary action into a 20-μm photopatterned alignment cell. The filled cell was then cooled to the desired temperature (phase) using an Instec HCS 402 heat stage. The filled cells were cooled at 2 °C/min to 70 °C, then 0.5 °C/min to 60 °C, and 0.25 °C/min at 0.5 °C increments thereafter. Samples were equilibrated at the designated temperature for at least 2 min. Thereafter, the mixtures were photopolymerized for 10 min with 365 nm light at an intensity of 50 mW/cmPhase transition temperatures and micrograph textures were obtained via polarized optical microscopy (POM) on a Nikon Eclipse Ci-POL in reflection with inline Instec HCS 402 heat stage. Kossel diagrams were captured on the same microscope with 100× oil-immersion objective (NA = 1.25), Bertrand lens, and Stress–strain measurements were performed on an RSA-G2 solids analyzer at a linear strain rate of 5% per minute. Glass transition temperatures were determined via differential scanning calorimetry (DSC) (TA Instruments Discovery DSC 2500). Transmission and reflection spectra of liquid crystal elastomers were collected with a Cary 7000 spectrophotometer (UV/Vis).Gel fractions were obtained by recording the mass of the polymer sample before and after washing with dichloromethane for 24 h.Supplementary informationDescription of Additional Supplementary FilesSupplementary Video


Listeria monocytogenes are extremely rare. Therefore, there is no standard procedure for their diagnosis and treatment. A 76-year-old Japanese man with diabetes and hypertension was diagnosed with a left common iliac aneurysm caused by L. monocytogenes, using multidetector computed tomographic angiography and rapid diagnostic testing of the positive blood culture. He was successfully treated with a combination of ampicillin administration, timely surgical debridement, and in-situ Y-graft placement with revascularization and omental implantation. Vancomycin and third-generation cephalosporins, to which L. monocytogenes is resistant, are used as an empirical regimen for infected aneurysms. Therefore, the use of a rapid diagnostic testing is important as it identifies L. monocytogenes within 24 h from obtaining the blood cultures, and guides the administration of the appropriate antibiotics. In-situ Y-graft placement restores nearly normal blood flow, following the confirmation of negative conversion of blood culture in response to the intensive intravenous ampicillin therapy. Appropriate and timely microbiological examinations, in addition to radiographic examinations, can be the key for selecting the optimal therapeutic procedures for each patient and achieving the best possible outcomes.Infected aneurysms caused by  Listeria monocytogenes is a food-borne pathogen, which causes rare but serious infections, particularly in the elderly, pregnant women, and newborn infants, owing to their compromised immune system . L.. L.L. mo at pH 4 . Proton fections . Our patL. monocytogenes infections; however, the optimal duration of preoperative antibiotic therapy varies, depending on several factors such as immunosuppressive comorbidities, urgency for rupture, and methods of the impending surgery . In Taiwan, two to six weeks of preoperative antibiotic therapy is considered optimal for treating infected aneurysms, unless urgent surgery is indicated [L. monocytogenes. We believe that a short course of preoperative treatment with appropriate antibiotics, under a condition of negative conversion of blood cultures, is acceptable for infected aneurysms caused by L. monocytogenes, which may cause prosthetic graft infections. In addition, the optimal duration of postoperative antibiotic therapy varies depending on pre-existing factors; the duration can range from nil to lifelong [L. monocytogenes affects prosthetic grafts and causes refractory prosthetic infections [L. monocytogenes infections. Our patient was disease-free with no exacerbation of graft infection after the 12-week antibiotic therapeutic course.Aminopenicillins are a group of first-line antibiotics for treating ndicated . Our patlifelong . A contilifelong . L. monofections . TherefoL. monocytogenes-infected aneurysms. A retrospective study did not show improved outcomes of aminoglycoside synergy with respect to early mortality (defined as death occurring 3–14 days after admission) [Clinicians need to make a choice between initial mono- and bi-antibiotic therapies for mission) . The premission) . We suspL. monocytogenes-infected aneurysms due to the high mortality rate ranging between 75 % and 100 %, resulting from aneurysmal rupture in patients undergoing only medical treatment for infected aneurysms [Surgical intervention is required in most cases of neurysms . In addineurysms . In our L. monocytogenes-infected aneurysms; therefore, the therapeutic approach needs to be tailored individually. Rapid diagnostic testing can be a guide for devising therapeutic strategies for emergent infections, such as L. monocytogenes-infected aneurysms. We believe that rapid diagnostic testing should be performed in cases where Gram-positive rods are observed in blood cultures, especially from immunocompromised patients with infected aneurysms. L. monocytogenes is associated with prosthetic graft infections; therefore, extra-anatomic reconstruction is indicated in most cases of L. monocytogenes-infected iliac aneurysms. However, in-situ reconstruction can also be proposed in cases where appropriate intensive antibiotic therapy is administered, as in our case. We believe that appropriate and timely microbiological examinations, in addition to radiographic examinations, aid in deciding the optimal surgical procedures for individual patients, which may ensure preferable outcomes.In conclusion, there is no universally accepted strategy for diagnosing and treating This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.This study has been approved by the institutional review board and ethics committee of Japanese Red Cross Ise Hospital .Written informed consent was obtained from the patient for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal on request.Hirokazu Toyoshima: Conceptualization, Methodology, Data curation, Writing - original draft, Writing - review & editing, Visualization. Koji Hirano: Methodology, Data curation. Motoaki Tanigawa: Supervision. Naoto Masuda: Conceptualization, Methodology. Chiaki Ishiguro: Conceptualization, Methodology, Supervision. Hiroyuki Tanaka: Methodology. Yuki Nakanishi: Methodology. Shigetoshi Sakabe: Supervision.The authors report no declarations of interest.


The prevalence of multiple HPV type infections was 32%. High-risk HPV types 52, 68, 35, 18 and 16 were the most frequent. A higher proportion of HIV-positive women had multiple HPV types compared with HIV-negative women. Conclusions: This study demonstrated a high prevalence of HPV in the study cohort. HIV-positive women were identified as having the highest HPV prevalence and infection with multiple HPV types across all ages. High-risk genotypes were the most commonly found.Background: Human papillomavirus (HPV) infection is now a well-established cause of cervical cancer and other anogenital cancers. An association between human immunodeficiency virus (HIV) infection and higher HPV incidence and prevalence are commonly reported. This study was conducted to demonstrate HPV prevalence, genotypes and its characteristics, according to the HIV status in women from Maputo in Mozambique. Methods: A total of 233 participants with ages ranging from fourteen to forty-five were included. Cervical samples were collected, DNA extracted, and HPV genotyping was performed using the HPV Direct Flow CHIP Kit. Results: In total, 177 HIV-negative and 56 HIV-positive women were included in the analysis. The overall HPV prevalence was 63% and was significantly higher among HIV-positive women (79% versus 58% among HIV-negative women;  Human papillomavirus (HPV) infection is now a well-established cause of cervical cancer and there is growing evidence of HPV being a relevant factor in other anogenital cancers  as well as head and neck cancers . CervicaBoth HIV and HPV are sexually transmitted viruses . Since H®, GlaxoSmithKline) targeting HPV 16 and 18, the quadrivalent vaccine , targeting HPV 6, 11, 16, 18 and nonavalent vaccine (Merck) which protects against quadrivalent vaccine types and 5 additional genotypes  [The HPV types 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73 and 82 are causally associated with vulvar, vaginal, cervical, anal, penile and oropharyngeal cancers  and more and 58) ,13. HPV  and 58) .The East African region is one of the most affected with an age standardized incidence rate of 42.7 per 100,000 women . In thisHIV infection and sexual debut before 18 years of age are factors associated with multiple HPV infections. HPV vaccine has not yet been introduced in the national expanded programme of immunisation in Mozambique. Taking into consideration the WHO global initiative to accelerate the elimination of cervical cancer by pursuing three important steps, namely: vaccination, screening and treatment, as well as the improve substantially cervical cancer outcome through early detection and effective treatment in low-income countries. To provide insights to inform the strategy for elimination of cervical cancer in Mozambique, the aims of this study was to determine the prevalence of HPV types and its characteristics stratified by age and HIV status, in women recruited from a Health Center (Maputo).This cross-sectional study was conducted on non-pregnant women seeking care regarding gynaecological symptoms such as: venereal pain, genital ulcers and vaginal discharge or even family planning in health facilities belonging to Mavalane Health’s area in Maputo, between February 2018 and July 2019. Exclusion criteria included pregnancy, current use of antibiotics or other antimicrobial medications , menstruation at the time of visit and vaginal douching during the last 7 days.The study followed the tenets of the Declaration of Helsinki of 2013. Written informed consent was sought from the participants. The Mozambican National Bioethics for Health Committee (CNBS) (ref: 423/CNBS/2018) and the Human Research Ethics Committees of the University of Cape Town (UCT) (HREC reference 850/2019), approved all aspects of the study. Sociodemographic data and information on risk factors were obtained through interviews.A total of 233 participants aged between 18 and 45 years old were recruited and the objectives of the study explained by members of the research team. After obtaining informed written consent, participants were interviewed twice using a semi-structured questionnaire regarding socio-demographical information, sexual behaviour and genital symptoms by different members of the research team in order to double-check answers. Pre-HIV testing counselling and an HIV-rapid test were performed in all women with unknown HIV serostatus, using the national serial testing algorithm which employs two sequential HIV-1/2 rapid tests: the Determine HIV 1/2 test , used for screening and UniGold HIV test , used to confirm initial reactivity on the Determine assay. Indeterminate samples were repeated, using fourth generation test Enzygnost ELISA Anti-HIV 1/2 Plus .After a speculum examination, cervical samples were collected by rotating a cytobrush at an angle of approximately 360° at the bottom of the posterior vaginal section. The brushes were placed in a test tube containing 10 mL of a SurePath liquid-based cytology  and stored at −80 °C. Two mL of this samples was transferred to the University of Cape Town (UCT) and used for deoxynucleotide acid (DNA) extraction and HPV genotyping.g for 30 min at 4 °C, and cell pellet was resuspended to 400 µL phosphate-buffer saline. DNA from resuspended cells was extracted by a MagNA Pure Compact  using the MagNA Pure Compact Nucleic Acid Isolation Kit  following the manufacturer’s instructions. HPV genotyping was performed using the HPV Direct Flow CHIP Kit  which allows the qualitative detection of 35 types of HPV  by amplification of a fragment in the viral region L1 of papillomavirus by PCR, followed by hybridization onto a membrane with DNA-specific probes by using the DNA-Flow technology for manual hybriSpot platforms . The biotinylated amplicons generated after the PCR were hybridized in membranes containing an array of specific probes in a three-dimensional porous environment for each target as well as amplification and hybridization control probes. Once the binding between the specific amplicons and their corresponding probes has occurred, the signal was visualized by an immunoenzymatic colorimetric reaction with Streptavidin−Phosphatase and a chromogen (NBT-BCIP) generating insoluble precipitates in the membrane in those positions in which there has been hybridization. The results were analysed automatically with the HybriSoft software .A total volume of 2 mL Thin Prep cervical specimen was centrifuged at 8000× p-values were <0.05.The data obtained through the questionnaire was checked for accuracy and double-entered into the Microsoft Excel (2016) computer programme. Exploratory analysis was first carried out to obtain descriptive statistics. Charts and tables were used to summarize data and display figures where appropriate. Data were analysed using Stata 14. HPV infection(s) were compared between HIV-negative and HIV-positive women, overall and stratified by age, using χ2 tests or Fisher’s exact tests in the case of sparse data. Trends in the prevalence of HPV by age were examined using non-parametric tests for trends across age groups. Single HPV infection was defined as infection where only one HPV type was detected, while multiple infection was defined by the presence of two or more HPV types. HPV was further categorised into high-risk (HR), low-risk (LR), or both high- and low-risk according to the types detected. Differences were considered to be statistically significant when p < 0.001; p = 0.088).A total of 177 HIV-negative and 56 HIV-positive women were included in the analysis, with the HIV-positive women being significantly older . This difference was most apparent among women aged 36–45 years: in this age group, the prevalence of HPV was 80% among HIV-positive women compared to 27% among HIV-negative women (p = 0.001). Among HIV-negative women, the HPV prevalence decreased with increasing age (p = 0.035). This scenario was not observed among HIV-positive women (p = 0.663).p = 0.001). These differences were most pronounced in the age groups 14–25 and 36–45 years. The prevalence of single HPV infection was 28% and did not differ by HIV-status (p = 0.832) or in any age groups.The overall prevalence of multiple HPV type infections was 32% but significantly higher among HIV-positive women .In the total sample, 28% of women had high-risk HPV only, 13% had low-risk HPV only, and 19% had both high- and low-risk types. The prevalence of high-risk only and low-risk only did not differ by HIV-status, but HIV-positive women were significantly more likely to have both high- and low-risk types , vaginal discharge (p = 0.852), or cervical inflammation (p = 0.426).No association was found between HPV and any of vaginal ulcer , 18 (7.7%), 17 (7.3%), 15 (6.4%) and 14 (6%) cases respectively. HPV-58 and 18 were the most frequent among HIV-positive women A. Among In p = 0.005); with a notably higher prevalence of multiple concurrent infections in the HIV-positive group. Several previous studies from the Sub-Saharan Africa region, identified a strong positive association between HIV and HPV infection [The study demonstrates a high HPV prevalence in this population 63%) which was significantly higher among HIV-positive women 79% versus 58% among HIV-negative women  in HIV positive women across the age groups of 14 to 25 years and 36 to 45 years while it generally decreased with age among HIV-negative women. In a study conducted by McDonald et al., (2014) similar results were seen among South African HIV-positive women  and theyIt is not biologically clear whether multiple concurrent HPV infections increase the risk of cervical cancer ,25. SomeIn our study, the most frequent HR-HPV types were HPV 52, 68, 35, 18 and 16. This is in partial agreement with other studies conducted in the same region. In a study conducted by Edna Omar et al. in Mozambican healthy young women, it was demonstrated that HR-HPV genotype distribution in Mozambican healthy young women differs from the global figures with HPV52, 35, 16, 53, 58 and 51 being the most prevalent .In the sub-Saharan African region, the most frequently detected HPV types in women with single and multiple HPV infections and normal cytology, were HPV 16 (50.7%), HPV 18 (19.2%), HPV 45 10.1%), HPV 35 (9.7%), HPV 33 (5.0%), and HPV 52 (4.5%) .1%, HPV . In our In the Eastern Africa regions where Mozambique is located, HPV-16 and HPV 52 are found being the most prevalent genotypes in women with cervical cancer as well as in the HIV co-infected women ,35. AlthA high prevalence of HPV, across all age groups, particularly in HIV-positive women was found in this cohort. This emphasises the need to introduce cervical cancer screening programs in this group of women who are at high risk of developing cervical cancer. The WHO established guidelines to consider routine screening and treatment of cervical pre-cancer and provided different strategies for a national screening program ,42. ThesUniversal access to vaccination is the key to avoid most cases of HPV-attributable cancers . AlthougThe study has limitations due to the lack of data available on cytology for the participants. In the HIV positive participants, we lacked data on anti-retroviral treatment.This study found a high prevalence of HPV in this cohort. Across all age groups, multiple HPV infection was higher in HIV-positive women compared to HIV-negative women. High-risk genotypes were the most commonly found in the study cohort. Stratifications in age and HIV status are advantageous in studying the prevalence of HPV in a cohort of sexually active women to define target populations for cervical cancer screening based on HPV and plan implementation of such programs. Overall, the study results highlight the need to intensify sexually transmitted infection prevention interventions but also the need to implement HPV vaccination and cervical screening programs, as recommended in the millennium development goals to reduce cervical cancer by 2030 .


Candida albicans in oral leukoplakia with different degrees of dysplasia.To identify human papillomavirus (HPV), Epstein–Barr virus (EBV), and C. albicans. To determine the association between each microorganism with different degrees of dysplasia a Chi‐square test was employed.An observational, cross‐sectional, descriptive study was performed using 30 formalin‐fixed and paraffin‐embedded tissues from patients with clinical suspicion of leukoplakia and confirmed diagnosis of oral dysplasia. Histological analyses were performed by two pathologists (interobserver) and dysplasias were classified as mild, moderate, or severe. Conventional PCR was used to detect HPV and EBV viruses and C. albicans (23.3%). Significant differences were observed between degrees of dysplasia and HPV presence (p = 0.005). In lesions positive for HPV, EBV, and C. albicans the most frequent histological changes were hyperkeratosis, irregular interpapillary ridges, and loss of basal stratum cell polarity.The tongue was the most common site for leukoplakias (71.4%) in females with a mean age of 50 years . EBV was the most frequently detected (73.3%), followed by HPV (43.3%), mainly of type 16 (40%), and Candida albicans in oral leukoplakia could be associated with dysplastic changes.Co‐infection with human papillomavirus, Epstein Barr virus, and  Still, a causal association between leukoplakia and EBV is yet unclear  in a total volume of 20 μl. Thermal cycling conditions were: 95° for 5 min, 95° for 1 min (1 cycle), 55° for 1 min, 72° for 1 min and 5 s (35 cycles), and one last cycle at 72° for 5 min. HeLa cells (positive for HPV) were used as a positive control and water as a negative control. Human papilloma virus PCR products were visualized in 3% w/v agarose gels in a Bio‐Rad Gel Doc XR+ documentation system. Each experiment was carried out in triplicate and internal controls were used.Specific primers were used to detect HPV in oral leukoplakias Table , under t2.42, and 1 μM of primers, 1.25 U GoTaq polymerase, 0.2 mM dNTPs, and 2 μl (0.5 ng) DNA sample in a final volume 20 μl. Thermal cycling conditions were: 95° for 5 min, 95° for 1 min (1 cycle), 60° for 1 min, 72° for 1 min and 5 s (35 cycles), and the last cycle at 72° for 5 min. An HPV16 plasmid was used as a positive control (donated by Doctor Zur Hausen of the Deutsches Krebsforschungszentrum—DKFZ of Germany), and water was used as a negative control. PCR products for HPV 16 were visualized in 3% wt/vol agarose gel in a Bio‐Rad Gel Doc XR+ documentation system. Each experiment was carried out in triplicate and internal controls were used.HPV‐positive samples were confirmed with specific primers for HPV16 . Thermal cycling conditions were: 95° for 5 min, 95° for 1 min, 65° for 1 min (1 cycle), 72° for 1 min and 5 s (35 cycles), and the last cycle at 72° for 5 min. An EBV sequence was cloned in a pUC57 plasmid vector and used as a positive control, and water was used as a negative control. PCR products for EBV 16 were visualized in 3% wt/vol agarose gels in a Bio‐Rad Gel Doc XR+ documentation system. Each experiment was carried out in triplicate and internal controls were used.EBV identification was carried out using specific primers for EBV Table  under th2.6C. albicans was identified by specific primers . The thermal cycling conditions were: 95° for 5 min, 95° for 1 min (1 cycle), 68° for 1 min, and 72° for 1 min and 5 s (35 cycles) and the last cycle at 72° for 5 min. These reactions were performed in a CFX96 Bio‐Rad device. C. albicans ATCC 90028 was used as a positive control, and water was used as a negative control. PCR products for C. albicans were visualized in 3% wt/vol agarose gels in a Bio‐Rad Gel Doc XR+ documentation system. Each experiment was carried out in triplicate and internal controls were used.rs Table , under t2.7C. albicans and different grades of dysplasia. A Chi‐square test was performed using the SPSS statistical software, version 2016, with a significance of p < 0.05.Data were analyzed using means, standard deviation, and table of frequencies to determine whether there was a relationship between HPV, EBV, and 33.1The study cohort consisted mainly of females (78.5%); with age ranging between 30 to 50 years old (57.1%). A higher incidence was observed in urban areas 64.2%), where the tongue was the most common site for leukoplakias  were positive for HPV (46% [n = 6] HPV16), 73.3% (n = 22) were positive for EBV, and 23.3% (n = 7) were positive for C. albicans  introduced the term “ epithelial precursor lesions” defined as epithelium alterations with a greater probability of progression to squamous cell carcinoma , similar to that reported in 2009 by Yang et al. who described a mean age of 48 years old (<65 years old) in patients with oral leukoplakia . However, others studies have described these clinical type in other sites of the oral cavity, such as gums (56%), palate (9%), lips (16.6%), tongue (25.7%), and the floor of the mouth . The main HPV serotype was 16, which was found in 40% of the samples included. Presence of these infections could lead to dysplasia in oral cavity tissues and induce malignant processes , followed by HPV (43.3%), in addition to p = 0.005) was observed between presence of HPV and cellular atypia at the level of dysplasia . All samples were collected from participants after signed informed consent under the ethical norms of the declaration of Helsinki.