Chlamydia trachomatis, HSV1, HSV2, Mycoplasma hominis and Ureaplasma urealyticum was analyzed. A prevalence of HPV infection of 27.4% was found. Interestingly, infections by exclusively low risk HPV genotypes or high/intermediate risk HPV genotypes were present in 64.5% and 22.6% of cases, respectively. Low risk-HPV6 was the most frequently detected genotype. Remarkably, HPV and C. trachomatis infections were significantly associated to each other . No significant differences in sperm quality were found between HPV-positive and HPV-negative patients indicating that HPV male urogenital infection does not impair sperm quality. Our results show a high prevalence of HPV urogenital infection among male partners of infertile couples, and that HPV and C. trachomatis infections are reciprocal risk factors of their co-infection. Moreover, our results suggest that men constitute a reservoir for continued transmission of C. trachomatis and HPV to women highlighting the need for routine screening for these two pathogens in male partners of infertile couples.The prevalence of HPV infection and its relationship with other sexually transmitted infections was analyzed in a cohort of 117 male partners of infertile couples from Cordoba, Argentina. Semen samples and urethral swabs were obtained and the infection with HPV,  Several conditions have been associated with male factor infertility such as abnormalities of urogenital tract, malignancies, endocrine disorders, immune conditions and sexually transmitted infections (STI). Nevertheless, infertility still remains idiopathic in an important proportion of men2.Sexually active couples who cannot achieve pregnancy after one year or more of regular unprotected sexual intercourse are currently considered infertile. Reported data indicate that approximately 10 to 20% of couples in reproductive age have fertility problems. Among the latter, male factor is involved in up to 50% of cases4. STI in men may cause urethritis, prostatitis, vesiculitis, epididymitis and orchitis4. Although manifest signs and symptoms are present in many STI, in others the course of the infection is asymptomatic leading to untreated chronic infections that could impair fertility4. Certainly, available evidence indicates a higher prevalence of sexually transmitted bacterial  and viral  infections in infertile men compared with fertile men6.STI have been proposed to induce male infertility through multiple pathophysiological mechanisms8. Low risk (LR) HPV genotypes can cause genital warts and respiratory papillomatosis, whereas persistent infection with high risk types (HR) can promote malignant transformations of epithelial cells in the cervix, but also in the vagina, vulva, anus, penis, mouth and throat10. The natural history of HPV infections in men is less known than in women11. Some studies suggest that male urogenital HPV infections usually clears spontaneously over time and only persists in a small fraction of the male population13. HPV may be detected anywhere in the male reproductive tract of infected patients such as external genitalia , anal region, perineum, testis, epididymis, vas deferens, urethra and semen14. HPV prevalence and load significantly varies in samples from different anatomic sites, showing higher proportions in external genitalia epithelial specimens than in semen16.Human Papillomavirus (HPV) is one of the most common sexually transmitted viral infections in males and females worldwide18. The impact of HPV infection on sperm quality is also a controversial matter21. On the one hand, some studies have indicated a significant association between seminal HPV infection and low sperm quality, mainly impairing sperm motility24, while other studies have found no association25. Interestingly, only a few of those studies analyzed the presence of other putative concomitant STI besides HPV infection that could also affect sperm quality25.Despite the increasing evidence of HPV prevalence in semen, the worldwide distribution of HPV genotypes and their risk for male infertility remain inconclusive. Two meta-analysis and systematic reviews of HPV infection in semen showed an overall higher prevalence in men undergoing initial evaluation and/or treatment for infertility) in comparison with men from general populationsChlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum and Herpes simplex virus were analyzed in this study.The aim of the present study was to evaluate the burden of HPV infection and its relationship with other STI in a cohort of male partners of infertile couples from Cordoba, Argentina. Prevalence and genotype distribution of HPV infection, sperm quality and the association of HPV infection with other STI such as 26. The study population age ranged from 25 to 63 years, while their female partner´s age ranged from 22 to 46 years. Primary and secondary infertility was present in 81.2% and 18.8% of cases, respectively, while the mean time being unable to get a clinical pregnancy was 2.3 years. No significant differences in the mean patient age or female partner age, type of infertility and the time being unable to get a clinical pregnancy in their female partners, were found between HPV + and HPV- male patients   Table  online; C. trachomatis, M. hominis, U. urealyticum, HSV1, HSV2 and common bacteria  infection within the total patient population and within HPV-positive and HPV-negative patients are shown in Table C. trachomatis (4/5), five M. hominis (5/24), seven U. urealyticum (7/28), three HSV1 (3/12), eight HSV2 (8/28) and two common bacterial infections  (2/17) were found in HPV positive men , while no significant differences were observed when comparing the presence of M. hominis, U. urealyticum, HSV1, HSV2 and other common bacteria between HPV + and HPV− group.On the one hand, when comparing frequencies of infection by C. trachomatis and HPV infections. On the contrary, other tested infections were not associated with HPV infection, with the following OR values: M. hominis , U. urealyticum , HSV1 , HSV2 , common bacteria  . The estimated odds ratios (OR) for HPV and different pathogens analyzed in the bivariate analysis are shown in Fig. 01. The eSperm quality from HPV-positive and HPV-negative patients was also analyzed. No significant differences were found in any of the sperm quality parameters  assessed between HPV-positive and HPV-negative patients Table . AlthougC. trachomatis infection in male partners of infertile couples.Results from the present study show that genital HPV infection is commonly present in a population of male partners of infertile couples from Argentina. However, no association between HPV infection and alterations in sperm quality was found. Nonetheless, and up to our knowledge, this is the first study that shows a significant association between HPV and 22. Analysis of the literature indicates a higher percentage of infection in infertile men compared with either the general population or fertile men18. The prevalence of HPV infection in male partners of infertile couples reported herein (27.4%) is slightly higher than the prevalence already reported in other regions or countries around the world27, with the exception of Golob et al., who reported a prevalence of 37% in the external genitalia and 13% in semen samples from male partners of infertile couples15. In Córdoba, Argentina an overall prevalence of 8.3% has been reported in semen samples from men from general population28. Thus, our results show higher prevalence of HPV genital tract infection in male partners of infertile couples with respect to men from the general population. The prevalence of male genital HPV infection appears to vary substantially between regions and countries, with the lowest reported in asian men29. In addition, the prevalence of genital HPV infection in men from Brazil, Mexico, and USA has also showed significant differences that could be explained by many factors such as diverse sexual behaviors and practices and, ethnic-specific factors in the frequency of germline variations in various immune regulating genes. All these findings highlight the importance to study the prevalence of STI in different parts of the world30. Regarding HPV genotype distribution, our results showed a strikingly elevated prevalence of HPV6, followed by HPV16, HPV18, HPV31 and HPV53. Although other authors found HPV16 as the most prevalent in semen samples31, our results agree with those published by Shigehara et al., who demonstrated that HPV6 was the most common HPV type in the urethral swabs obtained from men with urethritis32. It is important to take into account that our study used DNA extracted from a mixed sample composed of urethral swabs and semen to detect HPV. This approach enhances the possibility of detecting HPV infections in the lower and upper male genital tract, which could be related to the higher prevalence of HPV infection detected and to the particular HPV genotype distribution found. It has been reported that sampling different anatomical sites and/or using different combinations of samples increment the chances of detecting HPV infection since it often occurs at multiple anatomic sites within the same individual, being mainly detectable in the skin of external genitalia or in urethra swabs than in semen32. However, a main limitation of our study is that HPV detection was performed in combined specimens from different anatomical sites that were sampled together and not separately. Therefore, we could not assess the individual contribution of each sample to the total prevalence and possible differences in genotype distribution according to the sample analyzed. Nevertheless, bibliography indicate that different locations in the same patient often shares the same HPV genotype33. Indeed, when HPV genotypes were analyzed in penile skin, semen and vas deferens from the same patient, high concordance of specific HPV genotypes were found in penile skin and semen samples33. In addition, Golob et al. reported a higher prevalence of HPV infection in external genitalia than in semen specimens. Moreover, they found a high concordance in HPV types found in both samples15.The reported prevalence of HPV infection in men varies widely worldwide and it has been described to be 2–31% in the general population and 10–35.7% in men with unexplained infertility34. Some studies have shown that HPV male urogenital infection associates with reduced sperm motility35. On the contrary, and consistent with our findings, other reports have shown no association between HPV male urogenital infection and sperm quality alterations36. A possible explanation for this discrepancy may be the anatomical location of HPV infection. Using different experimental approaches, Foresta et al. showed that reduced sperm motility was found in HPV-infected patients. Interestingly, HPV attached to the head of spermatozoa was found in a significant fraction of these patients, suggesting that a direct interaction of HPV with spermatozoa could impair sperm motility35. Therefore, it is likely that reports in which impaired sperm motility was found in HPV-infected patients included patients in whom HPV infection is present in the upper and/or lower male urogenital tract, which would enhance the chances of HPV-ejaculated sperm interaction, rather than in external genitalia. Besides, it could be speculated that positivity in men could be a temporary phenomenon, not affecting semen quality. However, it seems that male infection is not short lived since follow-up testing to check HPV clearance indicated that 63.9% and 14.7% of infertile men were still HPV positive in semen after 12 and 24 months, respectively13. In addition, another important aspect that could have contributed to that controversial issue is the putative role of co-infections. Indeed, in most reports in which alterations in semen quality were shown in HPV infected patients, no other possible concomitant uropathogens infections were assessed, thus neglecting their possible contribution24.The relationship between HPV male urogenital infection and alterations in sperm quality is a matter of debateC. trachomatis are at higher risk of having abnormal cervical cytology37. Regarding C. trachomatis and HPV co-infection of the male genital tract, available evidence comes from studies focused on the potential oncogenic risk of these associated infections in specific groups of patients39. HPV co-infection with other STI, such as Neisseria gonorrhoeae, C. trachomatis, Mycoplasma spp. and Ureaplasma spp. has been previously described in men with urethritis32. STI prevalence reported in the present cohort of male partners of infertile couples were similar to those found in a recent study from our country40. Certainly, the prevalence of C. trachomatis and Ureaplasma spp urogenital infection in a large cohort of infertile patients were similar to that found herein. Nonetheless and up to our knowledge, we are reporting for the first time a tight association between HPV and C. trachomatis infections in male partners of infertile couples. Although this finding should be interpreted with caution because of low number of C. trachomatis positive cases found in our cohort, bivariate and multivariate analysis confirmed a significant association between HPV and C. trachomatis infections. This evidence highlights the importance of including the screening of urogenital infections in the initial diagnostic workup of the infertile couple.HPV co-infection with other STI was more thoroughly investigated in females. de Abreu et al. have shown that women with urogenital co-infection of HR-HPV with C. trachomatis, have been investigated as associated possible risk factors for HPV transmission and persistence cooperating in the carcinogenesis process and may be with fertility alterations, especially in the female genital tract. The effect of HPV and C. trachomatis co-infections was also analyzed in men with chronic prostatitis-related symptoms showing that men with HPV and C. trachomatis co-infection have reduced sperm motility and lower counts of sperm with normal morphological forms41. When we analyzed sperm quality parameters in patients co-infected with HPV and the other uropathogens under study, no significant differences were observed in sperm concentration, viability, motility or normal morphology with respect to patients infected with HPV alone. Unfortunately, sperm quality analysis in patients co-infected with HPV and C. trachomatis with respect to patients infected with C. trachomatis or HPV alone could not be performed due to the low number of cases of the sole C. trachomatis infection. It has been shown that both, HPV and C. trachomatis, can attach to human spermatozoa in vitro with a subsequent increase in sperm DNA damage42. Beyond dissimilar results regarding whether HPV presence in semen affects sperm quality or not, there is growing evidence showing that sub-fertile women receiving inseminations with HPV + semen show reduced clinical pregnancy rates43. Moreover, it has been shown that prophylactic therapy with HPV vaccination improves the reproductive outcome in infertile males with HPV semen infection44. Although no differences were found in the semen quality of patients studied herein, is possible that HPV and/or C. trachomatis transmission to the female partner could have consequences on the couple's fertility.Microorganisms causing chronic inflammatory diseases, such as C. trachomatis infections. Nevertheless, no significant semen quality alterations were observed in either patient infected with HPV alone or patients infected with HPV and other common uropathogens. However, since men provide a reservoir for continued transmission of C. trachomatis and HPV to women, our study highlights the need of including routine screening and monitoring of these urogenital infections in male partners of infertile couples.In conclusion, our findings revealed a high prevalence of HPV urogenital infection among male partners of infertile couples from Argentina, and a tight association between HPV and 45, primary infertility was diagnosed when the couple was unable to conceive a clinical pregnancy after at least 12 months of unprotected intercourse, while secondary infertility refers to couples that achieved a clinical pregnancy at least once in the past but are currently infertile.One hundred and seventeen (n = 117) male partners of couples seeking diagnosis for their infertility attending the Andrology and Reproduction Laboratory (LAR), Cordoba, Argentina were included in the study. Eligible men were aged 18 years or older that had semen analysis requested by their physician as part of initial fertility evaluation, after failing to conceive with their partner after one year of unprotected intercourse. Participants who met the following criteria were excluded: varicocele grades 3–4, cryptorchidism, antibiotic treatment within the previous three months and patients receiving chemo/radiotherapy. Following the worldwide-accepted WHO criteriag to obtain the pellet.The study was carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki) standards and the Argentinian legislation for protection of personal data . The experimental protocol was approved by the Ethics Committee and Internal Review Board (CIEIS) of the Oulton-Romagosa Medical Center, Cordoba, Argentina (code RePIS# 004). Participation in the study was voluntary and participants signed a written informed consent for their inclusion in it. A short questionnaire to inquire about their reproductive health history, their female partners and exposure to different risk factors for infertility was performed. Active smokers were defined as men consuming 1 or more packs of 10 cigarettes per day, whereas non-smokers were men that did not smoke. Semen samples and urethral swabs were obtained from each individual included in the study. Semen samples were collected after 3–5 days of sexual abstinence by masturbation and ejaculation directly into standard sterile containers and delivered to the laboratory within 1 h of collection. Urethral swabs samples were collected by health care professionals, using a Dacron collection swab  that was inserted 2 cm into the urethra and rotated for 360 degrees whilst removing it. The samples were placed in sterile tubes containing 1 ml of SPG  until processing. After removing an aliquot of semen to perform semen quality studies, the swab was placed in the paired semen container, mixed and centrifugated 10 min at 500 × 2, 25 mM of each primer and 5uL of extracted DNA for a final volume of 25 μL. Amplifications were performed in a PCR thermal cycler  with an initial denaturation step of 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 1 min, annealing at 55 °C for 1 min, extension for 1 min at 72 °C, and a final extension step of 5 min at 72 °C. PCR products were electrophoresed on a 1.5% agarose gel, stained with 1 mg/mL ECO-Gel 20,000 × Highway dye , at 95 V for 30 min in TBE buffer . A 100 bp marker  was used as a size standard. A DNA-free sample was used as a negative control and an HPV-positive sample as a positive control. The amplified DNA fragments were visualized on a transilluminator  with UV light. The samples showing specific amplification were further analyzed by HPV genotyping. Co-amplification of the human β-globin gene was performed as an internal control using the sense primer GH20 (5’ -GAAGAGCCAAGGACAGGTAC-3’) and the antisense primer PC04 (5’ -CAACTTCATCCACGTTCACC-3’) under the same amplification conditions as for HPV. HPV genotyping was performed by PCR-Restriction Fragment Length Polymorphism (PCR–RFLP) analysis as previously described47. In brief, PCR amplicons were subjected to digestion with 7 restriction enzymes  and subsequently were subjected to electrophoretic analysis on a 2% agarose gel at 60 V for 60 min in 1 × TBE buffer. A 100 bp marker  was used as a molecular size standard. After electrophoresis, the size of each fragment was determined. Genotypes were assigned after comparing the patterns of molecular weights of fragments for each HPV genotype47. A total of 71 different HPV genotypes with different associated oncogenic potential could be detected by this methodology: 36 low risk genotypes, 20 high risk genotypes, 10 intermediate risk genotypes (probably carcinogenic) and 5 genotypes of still unknown oncogenic potential.The detection of HPV and other uropathogens was performed by polymerase chain reaction (PCR) in DNA purified from the pellet obtained from each patient as described above. Total DNA was extracted using an AccuPrep Genomic DNA Extraction Kit  according to the manufacturer’s instructions and subsequently retaining the genomic DNA at 20 °C until processing. HPV was detected by PCR using the sense primer MY11 (5’- GCMCAGGGWCATAAYAATGG -3’) and the antisense primer MY09 (5’-CGTCCMAARGGAWACTGATC-3`), which amplify the L1 region of the viral genome. The reaction mixture consisted of 2.5 mM of each dNTP, 1 U of GoTaq DNA Polymerase (Promega Corp.), 0.6 mM of MgClC. trachomatis, HSV1, HSV2, M. hominis, U. urealyticum and other common bacteria was also performed in clinical samples from individuals enrolled in the study. The presence of Escherichia coli, Enterococcus faecalis, Proteus mirabilis and other common bacteria was analyzed by conventional bacteriologic culture  after 72 h incubation in 5% CO2 and aerobic conditions at 37 °C. The presence of M. hominis and U. urealyticum in semen samples was assessed by the commercially available MYCOFAST Screening RevolutioN colorimetric test  following the manufacturer's instructions. The presence of C. trachomatis, HSV1 and HSV2 was analyzed by PCR in total DNA purified from a mix of urethral swab and a fraction of the semen pellet from each patient, as detailed above. C. trachomatis was assessed by amplification of the ompA gene using the primers SeroA1, SeroA2, pCTM3, and of the cryptic plasmid gene using the CTP1 and CTP2 primers, as previously described48. To assess the presence of HSV1 and HSV2, a multiplex PCR with the primers CTGTGGTGTTTTTGGCATCA (sense) / GGTTGTGGAGGAGACGTTG (antisense) for HSV1 and CATGGGGCGTTTGACCTC (sense) / TACACAGTGATCGGGATGCT (antisense) for HSV2 was performed49.The search for 45, with some modifications. Sperm concentration, progressive motility (PR) and non-progressive motility (NPR) motility were assessed within 1 h after ejaculation. Sperm viability was analyzed using eosin Y (Sigma-Aldrich) staining. Sperm morphology was evaluated by the Papanicolaou technique and according to Kruger's strict criteria. The concentration of round cells was evaluated using the Makler counting chamber  and peroxidase-positive cells were quantified among round cells using a previously described cytochemical assay50. In brief, 20 µl of working solution (containing benzidine and H202) were mixed with 20 µl of liquefied ejaculate, incubated for 5 min at room temperature and counted at 400X magnification on a phase-contrast microscope50.Semen analysis was performed according to the WHO 2010 Semen Analysis Manual (fifth edition)https://www.graphpad.com/scientific-software/prism/) and IBM SPSS Statistics for Windows, version 25.0 . Data distribution was assessed by the Shapiro–Wilk test. Mann–Whitney or Kruskal–Wallis non-parametric tests were applied for statistical analysis followed by Dunn's multiple comparison test in the latter case. Data are shown as mean ± SD. For nominal data, Pearson's Chi-square test or Fisher's Exact Test, were first performed on a two-way contingency table (n = 117). Fisher's exact test was performed for small samples size. Subsequently, a bivariate and a multivariate logistic regression analysis was carried out to determine the associations between demographic, clinical and co-infection factors and HPV infection. Odds ratios (OR) measure with 95% confidence intervals (95% CIs) were calculated. The subjects with one or more missing data included in the analysis were excluded (n = 16), leaving an analysis group of 101 individuals. Differences were considered statistically significant when p < 0.05.Statistical analysis was performed using the GraphPad Prism version 7.0. (GraphPad Software, La Jolla, CA, USA Supplementary Information 1.Supplementary Information 2.


To determine the general and genotype-specific prevalence of HPV and to identify potential risk factors for the infection in a population-based screening of Peruvian women.A total of 524 samples were analyzed by PCR and a total of 100 HPV positive samples were found, of which 89 were high-risk, 19 were probably oncogenic, 9 were low-risk and 27 other HPV types. The 26–35 and 36–45 age groups showed the highest proportion of HPV positive samples with a total of 37% (37/100) and 30% (30/100), respectively. Moreover, high-risk HPV was found in 33.7% of both groups and probably oncogenic HPV in 52.6% and 31.6%, respectively. High-risk HPV were the most frequent types identified in the population studied, being HPV-52, HPV-31 and HPV-16 the most commonly detected with 17.6%, 15.7% y 12.9%, respectively. Demographic characteristics and habits were assessed in the studied population. A total of 62% high-risk HPV were detected in married/cohabiting women. Women with two children showed the highest proportion (33.8%) of high-risk HPV, followed by women with only one child (26.9%). Those women without history of abortion had a higher frequency of high-risk HPV (71.9%), followed by those with one abortion (25.8%).The online version contains supplementary material available at 10.1186/s13104-021-05588-7. The human papillomavirus (HPV) is a double stranded DNA virus, with worldwide distribution , 2. MoreIt has been reported that viral infections contribute to 15–20% of all human cancers, as several viruses facilitate the multistage development of cancers . HPV 16 Cervical screening and early treatment have successfully lowered cervical cancer incidence and mortality in developed countries, nonetheless, these measures have not been as effective in Peru , 10. HPVSince the recent introduction of molecular techniques for detection of HPV, it has become evident that screening with HPV tests protect better against future cancerous lesions than cytology-based screening alone, and consequently, virological screening programs are becoming increasingly recommended –14. It iHospital Regional Docente de Cajamarca, in northern Peru during September 2017 to July 2019. Asymptomatic women attending a gynecological outpatient health center, who had a history of at least 1 sexual encounter were studied. Exclusion criteria included: pregnancy, severe gynecological bleeding, previous hysterectomy, previous history of HPV-related disease including cancer, warts and cutaneous manifestations.A cross sectional study was carried out, using samples and data collected from a previous study performed in Samples were collected from the ectocervix and endocervix using a disposable cytobrush, they were stored in a tube containing phosphate buffered saline (pH:8.6) for preservation. Three aliquots of each sample were stored at − 20 °C until testing.Samples were kept at − 20 °C until analyzed. DNA extraction from 200 µL of cervico-vaginal samples was performed using the High Pure PCR Viral Nucleic Acid Kit , according to manufacturer’s instructions.HPV amplification was carried out using primers and conditions previously described . The HPVHospital Regional de Cajamarca, Peru. The samples were obtained in the Specialized Oncological Preventive Service of the Hospital Regional de Cajamarca, as part of the cervical cancer screening program. All samples were analyzed after a written informed consent was signed.This study was approved by the Research Ethics Board of the All analyses were processed with the IBM Statistical Package for the Social Sciences (SPSS) software version 21.0 .https://www.inei.gob.pe/).Maps were created using QGIS 3.12.3 software. Data for creating the map were acquired from the Instituto Nacional de Estadística e Informática  of high-risk HPV, followed by women with only one child 26.9%). Those women without history of abortion had a higher frequency of high-risk HPV (71.9%), followed by those with one abortion (25.8%). History of sexual abuse and extramarital affairs did not show a high frequency of HPV detection. Finally, 56% of women that underwent Pap test within one year or less were positive for HPV, with 44.9% presenting high-risk types  found in the study, which is higher than global pooled prevalence of 11% according to two meta-analyses .1% found. MoreoveRegarding specific genotypes, this study found that types 52, 16 and 31 were the most frequent high-risk HPV, with 17.6%, 15.7% and 12.9%, respectively. Conversely, type 18 was only found in one sample. According to literature, HPV-16 and HPV-18 are associated with approximately 70% of all cervical cancers . A systeHPV infection was also evaluated according to some characteristics of the population to identify potential predictors for infection. We could highlight the greater prevalence among married and cohabiting women, which differs from a study carried out by Castellague  that fouSexual behavior and habits are important determinants for HPV infection, as infection is most commonly acquired sexually. One of the most important factors that may have an effect on HPV transmission is condom use. A study carried out by Winer et al.  showed aAnother habit evaluated was the use of sexual toys, a previous study on university women showed that these devices are used in approximately 65% of sexual encounters , moreoveIn conclusion a high prevalence was found among Peruvian women, being the ones between 26–35 years-old and those married/cohabiting the most commonly affected. Also, it was observed a high frequency of high-risk HPV, being types 52, 16 and 31 the most commonly identified. This study highlights the importance to detect the general and genotype-specific prevalence in different populations, as these are important epidemiological tools. Screening with cytological and molecular methods are recommended particularly in women with risk factors.Firstly, an important limitation is that other sexually transmitted diseases were not evaluated. Also, general and specific genotype distribution of HPV varies greatly among geographic areas, therefore, the current information cannot be extrapolated to substantial different settings.Additional file 1: Table S1. Human papillomavirus types and oncogenic potential. Table S2. Most frequent HPV types identified in the population studied.


Human papillomavirus (HPV) is a common virus, and about 5% of all cancers worldwide is caused by persistent high-risk HPV infections. Here, we reported a comprehensive analysis of the molecular features for HPV-related cancer types using TCGA (The Cancer Genome Atlas) data with HPV status. We found that the HPV-positive cancer patients had a unique oncogenic process, tumor microenvironment, and drug response compared with HPV-negative patients. In addition, HPV improved overall survival for the four cancer types, namely, cervical squamous cell carcinoma (CESC), head and neck squamous cell carcinoma (HNSC), stomach adenocarcinoma (STAD), and uterine corpus endometrial carcinoma (UCEC). The stronger activity of cell-cycle pathways and lower driver gene mutation rates were observed in HPV-positive patients, which implied the different carcinogenic processes between HPV-positive and HPV-negative groups. The increased activities of immune cells and differences in metabolic pathways helped explain the heterogeneity of prognosis between the two groups. Furthermore, we constructed HPV prediction models for different cancers by the virus infection score (VIS) which was linearly correlated with HPV load and found that VIS was associated with drug response. Altogether, our study reveals that HPV-positive cancer patients have unique molecular characteristics which help the development of precision medicine in HPV-positive cancers. Human papillomavirus (HPV) is an important carcinogen since the HPV proteins E6 and E7 are intimately related to the events that cause malignant transformation of HPV-infected cells , 2. A glPD-1 immune checkpoint pathway in multiple cancer types . Survival analysis showed that HPV played an important role for patients’ prognosis. Furthermore, we analyzed the differences in the carcinogenic processes between HPV-positive and HPV-negative groups from three aspects: driver genes, genome instability, and mitotic carcinogenic pathways. The results implied that HPV might trigger cancer through the cell-cycle disorder rather than genome instability. The tumor microenvironment is significantly related to the improvement of cancer patient survival and treatment effect , 19. In http://xena.ucsc.edu/). The expression data were transcript per million (TPM) values with log2(x+0.001) transformed, and non-silent mutation was defined as gene-level mutation calls, where 1 represents non-silent mutation and 0 represents wild type. HPV expression  was collected from a previous study .External data with HPV status for model validation were obtained from the gene expression omnibus (GEO) with accession numbers GSE117973 and GSE151666. Cell line expression data and drug sensitivity data were downloaded from Genomics of Drug Sensitivity in Cancer  to select variables which have important impact on patients’ survival. Survival analysis was performed by “survival” package in R.We collected gene sets for DNA damage repair (DDR) pathways , mitoticOther genomic instability and immune indicators covering “loss of heterozygosity (LOH) frac altered,” “Aneuploidy Score”, “Copy Number Variation (CNV) burden”, “Microsatellite Instability (MSI) score”, “Mutation load”, “Cytolytic Activity (CYT) score”, “Cancer-testis Antigen (CTA) score”, “Neoantigens”, “B cell receptor repertoire (BCR)”, and “T cell receptor repertoire (TCR)” were obtained from previous studies , 28.In order to observe the spatial proximity of these samples, we put all samples to a two-dimensional coordinate system using Uniform Manifold Approximation and Projection (UMAP) dimensionality reduction through the “umap” package and then clustering analysis was performed by the “dbscan” package.-negative samples within a given cancer type was performed by the “DEseq2” package , “partial response” (PR), “stable disease” (SD), and “clinical progressive disease” (CPD). We explored the distribution of VIS in the four groups and calculated the proportion of chemotherapy response in different groups segmented by scaled VIS. GDSC data were divided into two categories according to the threshold of scaled VIS = 1, and the difference of the half maximal inhibitory concentration (IC50) was analyzed between the two categories.Fisher’s exact test was used to evaluate the difference of gene mutation frequency between HPV-positive and HPV-negative groups. All the comparisons of pathway activity and other indicators between the two groups were performed by the two-tailed Wilcoxon-sum rank test. In the GDSC data set, the difference of IC50 between two groups was calculated by the two-tailed T test or Mann–Whitney U test when the data were not normally distributed. All statistical analyses were performed by R.Clinically, HPV-positive patients in HNSC have a better overall survival than HPV-negative patients . To confTP53 in the HPV-positive group of CESC and HNSC indicated that their genomes were more stable. In the HPV-positive group, the lower mutation frequency of ARID1A in CESC as well as FAT1, CDKN2A, and FGFR3 in HNSC demonstrated that abnormal cell proliferation of HPV-positive patients did not arise from driver gene mutations. Although CYLD and ZNF750 mutations were enriched in HPV-positive HNSC, the samples with these two gene mutations together accounted for only 20% of HPV-positive ones. These results indicated that those genes in the HPV-positive group were not the same as TP53 which was the main cause of cancer in the HPV-negative group. Additionally, TP53 mutations in UCEC were enriched in the HPV-negative group, and PTEN mutations were enriched in the HPV-positive group. The total number of mutations with TP53 or PTEN exceeded 80% in both HPV-positive and HPV-negative patients. This result illustrated that there was no difference in the driver gene level between HPV-positive and HPV-negative groups in UCEC. We further applied the t-test to compare the expression levels of different mutation driver genes with the HPV-negative group in the three cancer types. As shown in CASP8, CDKN2A, FGFR3, and TP53 group was higher in HPV-positive than those in HPV-negative group in HNSC. The expression of PTEN and TP53 was higher in the HPV-positive group than those in the HPV-negative group in UCEC, but there were no differences in CESC.The mutation frequencies of several driver genes in the HPV-positive group with CESC and HNSC were significantly lower than those in the HPV-negative group  in the scaled VIS >2 group in TCGA data  attributes to p53-dependent enhanced proliferation in vitro and activates the FGFR pathway to accelerate tumorigenesis in vivo , E285K (3 of 28 mutations), and R273C/H/S (23 of 217 mutations) for HNSC, CESC, and UCEC, respectively. The R248 in p53’s DNA-binding domain (DBD) could interact with DNA’s minor groove directly and the R248Q mutation caused conformation alterations in areas of DBD far from the mutation site , because the immune infiltration and metabolism in TME are associated with patients’ prognosis –66. HPV otherapy Figure 3Since HPV has a significant impact on the tumor microenvironment which is crucial to the chemotherapy effect of cancer patients –69, we aAlthough there was a higher occurrence of HPV infection in males with STAD or HNSC : XY, JX, DX, XB, HW, YL, MC, WW, ZX, DZ, LC. Writing, review, and/or revision of the manuscript: XY, JX, DX, XB, HW, YL, MC, WW, ZX, DZ, LC. Administrative, technical, or material support : XY, JX, DX, KL, SZ. Study supervision: KL, JX, SZ. All authors contributed to the article and approved the submitted version.This work was supported by the key research and development project of Hainan Province ; the National Natural Science Foundation of China ; Hainan Provincial Natural Science Foundation of China ; the Open program for Key Laboratory of Tropical Translational Medicine of Ministry of Education  (2020TTM005); Major Science and Technology Program of Hainan Province ; Hainan Province Clinical Medical Center and Innovation Research Fund for Graduate Students .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.


Polymerase chain reaction (PCR) sequencing determined HPV types. Quantification of HPV16 E6 and E2 genes was performed to determine viral load and physical state. HPV DNA was detected in the cervix , blood  and paired peripheral blood and cervix samples . The most frequent types found in the cervix were HPV16, 18, 31, 33, 58, and 70, while HPV16, 18, 33, 58, and 66 were the most frequent types found in the blood. HPV DNA in the cervix was associated with previous sexually transmitted infections (STIs) , HPV DNA in the blood , and cervical lesions (CIN I/II or III) (p = 0.007). Binomial logistic regression showed that HPV DNA in the blood  and cervical lesions  were associated with HPV DNA in the cervix. However, we did not find an association between HPV DNA in the blood and cervical lesions (p = 0.385). Our results showed that only HPV DNA found in the cervix was associated with cervical lesions.This study evaluated the presence of Human Papillomavirus (HPV) DNA in the cervix and peripheral blood of women with cervical intraepithelial neoplasia  and healthy individuals. Overall, 139 paired peripheral blood and cervix samples of healthy women and women with CIN I, II, and III ( Cervical cancer is the fourth most common malignancy diagnosed in women worldwide, accounting for 6.6% of total cancer cases . Nearly It is well established that persistent infection caused by Human Papillomaviruses (HPV) is involved in cervical and head and neck carcinogenesis ,6,7,8,9.The paired cervix and blood samples were obtained from women during their gynecological consultations at Hospital das Clínicas, Pernambuco State, Northeast Brazil. The samples were collected from November 2010 to December 2016. A total of 139 women agreed and signed free and informed consent forms. This study was approved by the Ethics and Research Committee  and (CEP/CCS/UFPE under CAAE: 09307612.8.0000.5208). In this study, we examined 139 paired cervix and blood samples. Among these, 71 (51%) were from healthy individuals, 43 (31%) patients had cervical intraepithelial cervical grade I (CIN I), 16 (11.5%) patients had cervical intraepithelial cervical grade II (CIN II), and 9 (6.5%) patients had cervical intraepithelial cervical grade III (CIN III). Cervical cancer samples (paired cervix and blood samples) were excluded due to possible metastasis processes.DNA from cervical and peripheral blood was extracted using the DNeasy Blood & Tissue Kit , following the manufacturer’s instructions. Subsequently, all samples were quantified using a spectrophotometer (NanoVue Plus Spectrophotometer). DNA quality was confirmed by polymerase chain reaction (PCR), which amplifies a fragment of the β-globin gene with the primers described in The presence of HPV DNA was assessed by amplification with two sets of oligonucleotides. The first step was performed with the set of consensus and degenerate primers MY09/11 , which ahttp://blast.ncbi.nlm.nih.gov/Blast.cgi (accessed on 31 December 2016), was used to compare previously known HPV sequences.HPV-positive samples were genotyped by PCR followed by sequencing. All the HPV positive samples were sequenced by the dideoxy-terminal fluorescent method by using the ABI PRISM BigDyeTM Terminator Cycle Sequencing v 3.1 Ready Reaction kit . The sequences were analyzed using the Staden package  with the programs Gap4 (version 4.0) and Pregap4 (version 1.5), which were used to assemble the contigs of the sequences obtained from HPV DNA. After the contig assembly, the BLAST program, available at Real-time PCR (qPCR) was used to determine the viral load and physical status of HPV16-positive patients. For this, seven random paired cervix and blood samples derived from CIN I, II, and III patients and 12 samples derived from patients with normal cervical cytology were selected. HPV16 viral load was determined through E6 gene levels present in all patient groups. Each sample’s viral load was expressed as the number of copies of E6 in 50 ng of DNA . A seriep-values were two sided with a significance level of 0.05, except the Spearman´s rho and Kendall´s Tau_b correlations. All data were analyzed using SPSS version 26 .Clinical aspects of patients were compared between patients with HPV DNA-positive and HPV DNA-negative samples from the cervix by using chi-square and Fisher’s exact tests. We performed hierarchical binary logistic regression analysis to identify the variables associated with HPV positive in the cervix, HPV DNA in blood, and cervical lesions. The correlation of viral load of HPV16 DNA in paired cervical and blood samples was examined using the Spearman’s rho and Kendall´s Tau_b correlation coefficient tests. All p = 0.023; OR: 2.978; CI:1.34–7.821), HPV DNA in blood , and cervical lesions (p = 0.007)  of blood samples and 37.4% (52/139) of cervical samples. Patients’ socio-demographic characteristics, such as age, ethnicity, age at first sexual relation, number of pregnancies, contraceptive use, number of sexual partners, smoking, previous STIs, and cervical lesions, were compared according to HPV DNA positive and negative results in cervical samples. HPV DNA in the cervix was associated with previous STIs (= 0.007) .HPV DNA was detected in the cervix of 24% of healthy individuals, 48.8% of patients with CIN I, 56% with CIN II, and 55.5% with CIN III. In peripheral blood, HPV DNA was found in 46.4% of healthy individuals, 44.1% of patients with CIN I, 55.5% with CIN II, and 77.7% with CIN III. Regarding the cervix or peripheral blood samples, HPV DNA was detected in 46.4% of healthy individuals, 60% of patients with CIN I, 75% with CIN II, and 88.8% with CIN III. In paired cervix and peripheral blood samples, HPV DNA was found in 24% of healthy individuals, 32.5% of patients with CIN I, 38% with CIN II, and 44% with CIN III .The most frequently found genotypes in the cervix were HPV16, 18, 31, 33, 58, and 70, while HPV16, 18, 33, 58, and 66 were the most frequently found blood sample types. Globally, the genotypes most frequently found were HPV16 and HPV58 in both cervix and peripheral blood samples. Additionally, HPV16 was the most frequent genotype in the paired cervix and blood samples .p = 0.000; OR: 9.324; CI:3.612–24.072) and cervical lesions  were associated with HPV DNA in the cervix. In contrast, only HPV DNA in the cervix  was associated with HPV DNA in the blood (p = 0.00). In addition, we did not observe an association between HPV DNA in blood and cervical lesions (p = 0.385).Binomial logistic regression showed that HPV DNA in the blood  in blood and 0.2 to 138 (E6 copies/50 ng of DNA) in the cervix. A correlation was observed between HPV16 viral load present in the cervix and peripheral blood samples in patients with CIN II/III (r = 0.65) and CIN III (r = 0.60) We did not observe a correlation between HPV16 viral load in the cervix and peripheral blood samples in patients with CIN II/III (r = 0.25) and CIN III (r = 0.20) . AdditioRegarding the healthy individuals, the viral load varied from 2.76 to 83.89 (E6 copies/50 ng of DNA) in blood and from 0.12 to 246 (E6 copies/50 ng of DNA) in the cervix. The integrated, episomal, and mixed physical status were observed in peripheral blood and cervix samples. However, we did not observe a correlation between HPV DNA viral load in the cervix and peripheral blood samples in healthy patients (r = −0.15).This study investigated the presence of HPV DNA in paired peripheral blood and cervix samples from women with cervical lesions and from healthy individuals. Our results showed HPV DNA in the cervix and peripheral blood samples of both healthy individuals and CIN patients. In addition, we also observed a significant association between the HPV DNA in the cervix and cervical lesions. However, we did not observe an association between HPV DNA in blood and cervical lesions. Moreover, we did not find a positive correlation between HPV16 viral load in the paired cervix and blood samples in women according to CIN grade and physical status of the virus in the cervix and blood. Globally, our results showed the presence of HPV DNA in the cervix and peripheral blood from cervical lesions patients and healthy individuals. However, only HPV DNA found in the cervix was associated with cervical lesions.Several studies have demonstrated the presence of HPV DNA in blood from cervical cancer ,34,35,36In this study, we also observed a direct relationship between the increase in prevalence of HPV DNA in blood and the severity of the lesions in patients with CIN I (44.1%), II (50%), and III (77.7%). Similar results were observed in previous studies regarding patients with cervical lesions . HoweverTo date, few studies discuss the presence of HPV DNA in the blood of healthy subjects ,24,32. HOne of the HPV-induced process’s key events is integrating the virus’s genome into the host’s genome . The sta+, and CD56+ [HPV DNA in peripheral blood has been historically detected in cervical cancer and non-cervical cancer tumors, in which ctHPVDNA migrates from local infection to blood . Furthernd CD56+ ,24.In conclusion, we detected HPV DNA in the cervix and peripheral blood from patients with cervical lesions and from healthy individuals. However, only HPV DNA found in the cervix was associated with cervical lesions. More studies are needed to clarify the role of HPV DNA in blood and tumorigenesis.


With the proliferation of live streaming, there is evidence that online impulse buying is becoming an emerging phenomenon. Although many studies have investigated impulse buying in the context of offline shopping and business-to-consumer e-commerce, online impulse buying in live streaming has attracted little attention. In this study, we aim to explore the effect of social presence in live streaming on customer impulse buying based on the stimulus–organism–response framework. The research model presented here identifies pleasure and arousal as the mediation of impulse buying in live streaming. We use the AMOST and IBM SPSS PROCESS software to estimate our model based on data at the minute level from 189 customers, who watched live streaming in the past three months. The results suggest that the social presence of the broadcaster and the social presence of the live streamer positively affect impulse buying directly and indirectly via pleasure and arousal, promoting consumer online impulse buying in live streaming, but the social presence of the viewers has no significant effect on pleasure and arousal. For practice, our results can help policymakers and operators of the live streaming platform alleviate impulse buying in the digital world. The prevalence of live streaming has driven a boom in e-commerce activities, namely live streaming e-commerce ,2,3,4. TIn typical live streaming e-commerce, the broadcaster can create and deliver real-time video to the customers. The broadcaster can engage potential audiences to join the live stream to consume the content. For example, the broadcasters can talk about the last beauty trend, introduce the product, and try on different items for the viewers. During the content consumption process, the viewers can also ask questions about the products, chat with the broadcaster and other viewers via text, and even send a virtual gift to their appreciated broadcasters. Compared with the traditional web-based e-commerce or social e-commerce, live streaming e-commerce can not only allow the customer to become closer to the products but also to hear the broadcaster describe how the product feels and watch the effect of the broadcaster try it on, thus fostering a more authentic and interactive online shopping experience ,10. ManyImpulse buying refers to an unplanned purchase, the result of exposure to a stimulus, and deciding to purchase on the spot ,12. AccoDuring the last decade, adequate research attention has been paid to online impulse buying. Current findings in the context of web-based e-commerce and social e-commerce context may not be suitable for impulse buying in live streaming e-commerce ,17,18. CTo address this question, we use the stimulus–organism–response (SOR) framework to conceptualize our theoretical model because the SOR framework provides a suitable lens to understanding the mechanism for impulse buying via social presence in live streaming. The SOR framework of environmental psychology was proposed by Mehrabian and Russell . The SORBased on the SOR framework, we construct and empirically test a structural model that examines impulse buying as a behavioral response caused by the social presence in live streaming e-commerce by using sampling data of 189 consumers from Taobao Live in China. Our study not only contributes to the literature on live streaming e-commerce and social presence theory but also provides practical insights for policymakers and live streaming operators to alleviate impulse buying in live streaming e-commerce.The remainder of the paper is organized as follows. First, we review three streams of related literature. Second, based on current studies, we propose a theoretical model and our hypotheses. Third, we introduce our research context, sampling and data collection, and measure items. Fourth, we empirically test the research model and hypotheses, followed by a presentation of the research results. Finally, this paper concludes with the theoretical and practical implications and limitations that give rise to further research. Live streaming is a type of user-generated content . It alloLive streaming e-commerce is more likely to induce customers’ impulse buying behaviors than traditional website e-commerce for the following reasons. First, in the traditional e-commerce website, interactive activities mainly occur between the customer and the website features . HoweverWith the development of live streaming e-commerce, growing attention has been paid to this emerging topic. However, the current research efforts have mainly focused on customer engagement and purchase intention in live streaming e-commerce ,10,30; iOnline impulse buying can be defined as “a purchase that is unplanned, the result of an exposure to a stimulus, and decided on the spot” . Some reWith the progress of live streaming e-commerce and its application, online impulse buying is common among customers ,35,36. FSocial presence refers to the “degree of salience of the other person in the interaction and the consequent salience of the interpersonal relationships” . The conSocial presence plays an important role in the online shopping context. Currently, social presence has often measured the warmth of media or human feeling of sociability from ICT . HoweverThe social presence of the broadcaster refers to the extent to which customers perceive the direct interaction with the broadcaster in live streaming ,24. The The social presence of viewers refers to the extent to which customers perceive the presence of other customers in the live streaming . The chaThe social presence of live streaming refers to the live streaming’s capability to convey a feeling of human contact, sociability, warmness, and sensitivity . In tradMehrabian and Russell  indicateOur research model is shown in In live streaming e-commerce, the broadcaster can introduce and try the products as well as provide personalized services for the audiences . For exaHypothesis 1a (H1a).The social presence of the broadcaster has a positive influence on customers’ arousal in live streaming e-commerce.Hypothesis 1b (H1b).The social presence of the broadcaster has a positive influence on customers’ pleasure in live streaming e-commerce.In live streaming e-commerce, the broadcaster presents various content to interact with the existing customers and attract new viewers into the virtual showroom. The audiences in the live streaming can also interact with the other viewers using text in the chat channel . CommuniHypothesis 2a (H2a).The social presence of viewers has a positive effect on customer arousal in live streaming e-commerce.Hypothesis 2b (H2b).The social presence of viewers has a positive effect on customer pleasure in live streaming e-commerce.The social presence of live streaming assesses the subjective characteristics of live streaming e-commerce, which reflect the social side of IT-enabled communication. Prior empirical evidence also suggests that the social presence of media communication can enhance the emotions of pleasure and arousal . Some reCompared with online e-commerce websites, live streaming e-commerce has more human touch and sociability with the broadcasters and other viewers ,5. Thus,Hypothesis 3a (H3a).The social presence of live streaming has a positive influence on customer arousal in live streaming e-commerce.Hypothesis 3b (H3b).The social presence of live streaming has a positive influence on customer pleasure in live streaming e-commerce.According to Mehrabian and Russell’s  model, tHypothesis 4 (H4).Consumer arousal is positively related to impulsive buying in live streaming e-commerce.Hypothesis 5 (H5).Consumer pleasure is positively related to impulsive buying in live streaming e-commerce.In this research, we considered Taobao Live, one of the major e-commerce platforms run by Alibaba in China, as our research context. Taobao Live is the live streaming service integrated into Taobao’s e-commerce App. In 2018, Taobao Live provided over 3000 live broadcasts under the guidance of its live streamers [In typical live streaming e-commerce, broadcasters can build the video streams and display and introduce the products  for customers. Customers can interact with the broadcasters and other customers using text. www.wjx.cn, accessed on 17 February 2022) to develop the questionnaire and chose a professional data collection service offered by the website to collect our data. Wenjuanxing has access to a sample database of 26 million Chinese respondents [An online survey was conducted to collect data to test our hypotheses. Specifically, we used the Wenjuanxing website  and late  waves of the completed surveys. The Mann–Whitney test shows that the two groups do not differ in age (p = 0.81), gender (p = 0.69), or income (p = 0.52). These results suggest that the nonresponse bias is not a concern in this study.4.3. Measure ItemsThe multiple-item scales used in our study are designed based on the relevant literature, and minor changes were made to fit the context of live streaming e-commerce see . Social The demographic information of surveyed respondents is shown in As our data were collected from a cross-sectional survey, common method bias (CMB) may exist. Following , we usedTo examine the reliability and validity of measures in our model, we conducted three types of necessary tests: internal consistency, convergent validity, and discriminant validity. First, we used the composite reliabilities  to asses2 = 0.703). The presence of the broadcaster, the presence of the viewer, and the presence of the live streaming proved to be effective in creating customers’ pleasure (R2 = 0.518) and arousal (R2 = 0.658). All these results provide strong support for our structure model.We used AMOST to estimate the path coefficient and the significance of the hypothesized relationship. In AMOST, we used the fit criterion of χ²/df, GFI, RMR, CFI, NFI, and NNFI to evaluate the structure model. Most of the criterions exceed the stipulated threshold except for PNFI: χ²/df = 1.197, GFI = 0.924, RMSEA= 0.032, RMR = 0.031, CFI= 0.984, NFI = 0.913, AGFI = 0.895, and NNFI = 0.981. AGFI is below its threshold (0.9), but it is common that not all fit criteria are perfect in the structural model . The strp < 0.01) and pleasure . These findings support H1a and H1b. SPOV has no significant effect on arousal  and pleasure . As a result, the findings do not support H2a and H2b. SPOLS has a direct and positive relationship on arousal  and pleasure . These findings support H3a and H3b. The coefficients of social presence of live streaming on both arousal and pleasure are stronger than the coefficients of social presence of the broadcaster on arousal and pleasure. The estimated path coefficients, related hypotheses, and variances explained are shown in p < 0.01). Pleasure has a direct and positive relationship on IB . These findings support H4 and H5. These results are consistent with prior studies; pleasure and arousal were the critical determinants of impulse buying [Arousal has a direct and positive relationship on IB  impulse buying in live streaming e-commerce. We used PROCESS in SPSS ,60 to coThe objective of this study was to understand consumer impulse buying in live streaming. Using the SOR framework as our theoretical lens, we investigated the role of social presence (stimulus) in impulse buying (response) in live streaming e-commerce by considering customer pleasure and arousal (organism). The findings confirm that the social presence of the broadcaster and the social presence of the broadcaster of the live streaming can predict customer arousal and pleasure. However, our findings suggested that there is no direct effect of the social presence of the viewers on impulse buying in live streaming e-commerce. One plausible explanation is that customers may suspect the motives of revealing the social cues or the authenticity of those social cues from other viewers; thus, these social cues from other viewers are not used by the customer to make purchase decisions . AnotherFurthermore, consistent with the current literature in both relational marketing and information systems ,31, impuThe study provides several theoretical implications. First, we contribute to the literature on customers’ well-being in the pandemic period by investigating the impulse buying behaviors in the context of live streaming e-commerce. Impulse buying behaviors are commonplace in live streaming e-commerce all over the world. Moser et al.  indicateSecond, this study contributes to the social presence theory by identifying a three-dimensional social presence in live streaming e-commerce: the social presence of broadcaster, the social presence of viewers, and the social presence of live streaming. Impulse buying in live streaming is typical IT-related behavior, and social presence is one of the critical design principles for customers’ IT-related behavior. The prior studies mainly examined the role of one or two dimensions of social presence  in IT-related behaviors ,41,42. IThird, our study sheds light on the formation mechanism of impulse buying in live streaming e-commerce by investigating the customer’s emotional state as a mediator. Pleasure and arousal (as a set of emotions) as two key mediating variables have been fruitful in explaining experiential shopping behaviors. Prior studies focused on the effect of specific stimuli such as website quality on customer impulse buying ,35,36. IThis study also presents important practical implications. First, in the context of COVID-19 emergence, investment in live streaming e-commerce has grown rapidly . Live stSecond, although the social presence of viewers seems to be a weaker prediction for customer pleasure and arousal, this does not mean that viewer presence does not matter for online impulse buying in live streaming. Policymakers and operators of the live streaming platform can limit the number of viewers in the live stream to reduce the social presence of other viewers. Conversely, policymakers and operators should pay more managerial effort to prevent these capper consumers from spreading false product information during live streaming.Third, the findings show that the customers’ pleasure and arousal play a mediate role between the three dimensions of social presence and impulse buying in live streaming. Therefore, a practical implication for operators of the live streaming platform is to design live streaming features  to alleviate customers’ excited emotional state when they are shopping online. At the same time, consumers should warn themselves not to engage in live streaming e-commerce when they are in a pleasant or stimulated emotional state.In conclusion, the main contributions of the study are three-fold. First, during COVID-19, the offline commerce operating modes were seriously interrupted by the emergency. Live streaming has empowered vendors to reach a large audience and to promote their products virtually face-to-face with interested buyers, leading to serious online impulse buying behavior. Based on the SOR framework, we attempted to understand customer impulse buying in live streaming, a relatively new e-commerce phenomenon. Second, we proposed a multi-dimensional conceptualization of social presence in live streaming to overcome the limitation of the one-dimensional social presence construct. Third, we highlighted the importance of three social presence factors in live streaming e-commerce that induce impulse buying via customers’ emotional state . This study has some limitations. First, the research context is Chinese live streaming e-commerce, which may restrict the external validity of our research findings regarding cultural differences. Future studies can further consider a sample of live streaming users in other countries to examine the external validity of our findings. Second, we are not restricted to a specific market  in live streaming e-commerce. It is an empirical question to whether our findings are replicable in a different market. Future studies should modify and apply the current research model to a specific product market. Third, in our research, we focused on social presence in live streaming to conceptualize customers’ virtual experience, which is relevant to inducing impulse buying. Future studies can extend our multi-dimensional construct of social presence that could be induced by live streaming to examine how different constructs may influence impulse buying in different ways. Finally, there are different modes of live streaming e-commerce: live streaming embedded in e-commerce  and e-commerce embedded in live streaming . Future studies can investigate the effect of social presence on impulse buying in the two different modes of live streaming e-commerce.


We consider the opportunities and challenges associated with organizing a conference online, using a case study of a medium-sized (approx. 400 participants) international conference held virtually in August 2020. In addition, we present quantifiable evidence of the participants' experience using the results from an online post-conference questionnaire. Although the virtual meeting was not able to replicate the in-person experience in some aspects (e.g. less engagement between participants) the overwhelming majority of respondents found the meeting an enjoyable experience and would join similar events again. Notably, there was a strong desire for future in-person meetings to have at least some online component. Online attendance by lower-income researchers was higher compared with a past, similar-themed in-person meeting held in a high-income nation, but comparable to one held in an upper-middle-income nation. This indicates that online conferences are not a panacea for diversity and inclusivity, and that holding in-person meetings in developing economies can be at least as effective. Given that it is now relatively easy to stream contents of meetings online using low-cost methods, there are clear benefits in making all presented content accessible online, as well as organizing online networking events for those unable to attend in person. Conferences are an essential part of any academic career, and offer a unique opportunity for scientists to interact, network and form partnerships. The absence of conferences can have greater impacts on early-career researchers  because they have time-sensitive academic paths that often rely on conferences to disseminate their work, connect with established researchers and identify future opportunities to advance their academic careers.During 2020–2021 some meetings proceeded on their original timetable yet switched to fully virtual or allowed virtual attendance (online-accessible) , while cHere, we provide some practical considerations for prospective organizers of online conferences. We also present a case study of a recent, medium-sized (approx. 400 participants), online international meeting, provide information on logistics and demographics, and analyse participants' feedback drawn from a post-meeting questionnaire.. 2The points below summarize general planning issues. (a)Live presentations are more interactive, but carry more risk , require technical support and place demands on presenters from different time zones. A pre-recorded format only depends on technical support, but is potentially less interactive. Offering both formats is more flexible but requires more planning. (b)This allows participants from other time zones, with a poor Internet connection, and with other duties to watch at their convenience. A logistically easy option is to use software (e.g. Zoom) that allows for live broadcasting on streaming services (e.g. YouTube) that offer automatic archiving of the streamed content by default. This content can be made publicly available or restricted to specific users as appropriate. If another platform is used that does not have the built-in capability to stream content on demand it may be logistically more complex to achieve, as it requires recording all live presentations, and uploading and hosting all presentations on the platform for the determined archival period. If presentations are made available on demand, consideration should be given to uploading standard instead of high definition videos, to minimize costs for those participants that need to buy Internet data packages. Another point is to add time stamps for the start of each presentation in the video description on YouTube, which enables the viewer to jump to specific talks by clicking on the time stamp that YouTube automatically hyperlinks to the spot in the video. Alternatively, individual videos of each presentation can be produced and uploaded, which takes more time but also results in a much more valuable product for the presenter. (c)https://2020.carpentrycon.org; GeoHab2021, http://geohab.org/geohab-2021; Virtual Island Summit 2019/2020/2021, https://islandinnovation.co/virtual-island-summit-2021).Identifying time zone representation of potential participants  can aid this decision. For hybrid meetings, the time zone is typically the same as the host institute, whereas for fully virtual meetings the time zone can be chosen to maximize participation and/or diversity. For international conferences, it is not possible to accommodate all participants' time zones, hence an on-demand option or splitting sessions across time zones might be appropriate Fewer (e.g. two to three) longer 10–12 h) days allow more individuals from different time zones to attend the meeting, although remaining focused for long periods can be challenging [ h days a (e)Platform selection will depend on presentation format , audience size, other services and events  and budget. A wide variety of platforms and relevant software are available, each with their own strengths and limitations . Thus, o (f)https://support.zoom.us/hc/en-us/articles/203806119-Restricted-countries-or-regions) is important. It is essential to define a code of conduct, which should at its minimum outline examples of acceptable and unacceptable behaviours, as well as consequences for those violating the code. An example code of conduct can be found in electronic supplementary material, S1. All of the above should be clearly communicated with participants in advance so that they are confident that due diligence has been taken to make the online meeting environment a protected and safe space.Conferences are often a venue for presenting unpublished results, so it is important to take measures to ensure that information will not be recorded or posted on social media without consent. The security of platforms and whether particular platforms are banned from being used in specific sectors and/or countries  .. 3https://dsbsoc.org/). The meeting focussed on early career researchers for whom conferences are critical career milestones, and while opportunities to present at meetings during the COVID-19 pandemic were significantly reduced.eDSBS was an online-only meeting held 19–21 August 2020, and organized by a UK-registered charity, the Deep-Sea Biology Society ( (a)https://zoom.us/webinar) was used for live-streaming of oral presentations, and presenters could choose to present live or have a pre-recorded talk streamed by technical support during their assigned session . This enabled live questions and discussion similar to an in-person conference. All talks were recorded and subsequently uploaded on the main meeting website , where they were available for 14 days after the live presentation. Therefore, regardless of participant time zone or stability of their Internet connection, participants could login to the main meeting website, open the sessions, go through the recorded talks and participate in text-based discussions in their own time. Poster presentations were not presented live, but all posters were available in poster halls on the main meeting website for the duration of the conference and the subsequent 14 days. Each poster was supported by text-based discussion and highlighted in an online poster browsing session (i.e. slide show streamed via Zoom Webinar). Finally, social events  were held in Zoom Webinar, where occasionally participants were randomly assigned to breakout rooms to maximize interactions and networking opportunities.Prior to the meeting, a questionnaire was sent to the Society membership to gauge interest and help inform decisions on the meeting format . Upon feedback, the organizing committee settled on an online format that was semi-synchronous with live-streaming as well as on-demand oral and poster presentations. The meeting observed a strict code of conduct  to ensure participants were comfortable sharing slides and posters. Only logged-in and registered participants were able to access presentations, and it was not possible to download videos.https://slack.com) was used for further participant engagement, troubleshooting issues, and rapid communication of announcements and communication between organizers. Both email and Slack were used for troubleshooting and support for all the participants. Finally, Google Workspace (https://workspace.google.com/) was used to store information on logistics, data cloud, forms and to facilitate email communications. Python scripts for converting forms into abstracts and timetables are available at https://bitbucket.org/beroe/conference-generator.Numerous methods were used to communicate and share information with meeting participants. The Society website contained all essential information related to the meeting such as registration and abstract submission deadlines, schedule of talks, book of abstracts and others. Social media (e.g. Twitter) was used to advertise the event and associated deadlines, to engage a wider audience and to post scientific content during the meeting. Pre-meeting announcements, including instructions on how to access the main meeting website, links for all Zoom sessions and guidelines for presenters and session chairs, were sent via email. Slack ( (b)The conference organizers sent a questionnaire to participants at the end of the meeting requesting participants to rate statements  related to different aspects of the meeting . Participants regarded live talks as a key component of online meetings , although the total number of questions they received and overall engagement with other researchers was deemed less than in-person meetings . Less engagement and social interaction with other researchers has been consistently identified by organizers and participants alike as a major disadvantage of virtual meetings e.g. ,17,18). ,18. 17,1Overall, the overwhelming majority of participants found the online meeting an enjoyable experience and would join similar events again 94%, Q52). Similar sentiments have been reported for online conferences either anecdotally (e.g. ) or by p4%, Q52. All participants' responses are available in electronic supplementary material, S4. (c)Additional feedback covering logistical, technical and practical aspects of organizing a meeting is available at electronic supplementary material, S5. (d)a). This was achieved by the scientists themselves using ‘off-the-shelf’ software products and no commercial conference organization was hired. For virtual participants, the registration cost was considerably lower (approx. 90–97% less) compared with in-person meetings (b), although the costs were heavily subsidized from a grant and also those organizing the meeting did not charge for their time . Overall attendance cost was even lower though, as virtual meetings do not entail travel, accommodation and subsistence costs that in-person meetings do, and which can be prohibitively expensive for many researchers.The cost of conducting the online eDSBS was only 4% of the cost of in-person meetings with similar themes and levels of attendance a. This wmeetings b, althouc), and a 50% increase in the number of low and middle-income countries represented in the meeting (d). The relative numbers of participants from low and middle-income countries was a significant improvement (double) compared with the USA meeting . The figures from eDSBS appear to be a step in the right direction. While 14% might still seem low, deep-sea biology is in general poorly represented in low and middle-income countries [In an effort to encourage participation and increase inclusivity, eDSBS waived registration fees for participants from developing economies and in difficult financial situations. Comparing demographic composition to a similar-themed, past in-person meeting held in the USA, a high-income country, eDSBS had almost twice as many participants from low and middle-income countries c, and a  meeting d. The reountries  owing toc,d). Notably, the relative numbers of participants from low and middle-income countries was actually higher for the Colombia meeting , a result of increased participation of researchers from the host and neighbouring countries. This is a notable finding, suggesting online conferences are not a panacea to combat reduced participation from lower- and middle-income researchers, and that holding in-person meetings in developing economies can be at least as effective. It is also likely that if greater funds are available to bring researchers from low and middle-income countries to in-person meetings this would also help redress this balance, without needing to go online-only.When comparing eDSBS demographics to a past in-person meeting held in Colombia—an upper-middle-income country—the number of participants from low and middle-income countries, as well as the total number of countries represented in the meeting, was comparable c,d. Nota meeting c,d, a rec,e) was due to a combination of factors such as the meeting's particular focus on this group, the virtual format and ease of use, as well as the much lower registration costs compared with in-person meetings which has led to enhanced early-career participation elsewhere too . The. Thec,e). 4ad hoc meetings, chats over coffee and casual dinner invitations, etc. that a week-long in-person conference creates, many of which may be critical to a person's career. It is important that conference organizers remain committed to providing the in-person interactions that are so essential to early-career researchers, and that the supporters of those young persons recognize the need to fund those events. At the same time, given the large carbon footprint that in-person meetings can carry [While it is possible to directly gain useful feedback on the successes and failures of online meetings such as the results presented here, it is not straightforward to quantify the networking and information flow in an online meeting compared with an in-person meeting. For example, it is difficult to survey all of the in-person interactions, an carry , it is iClick here for additional data file.