This paper examines the effects of customer personality, trust, and satisfaction on customer loyalty in premium banking services. Based on a survey of 210 high-net-worth premium banking customers, the study found that the customer personality dimensions of conscientiousness, extroversion, and agreeableness affected trust, while extroversion affected satisfaction. Trust displayed a significant influence on both satisfaction and customer loyalty, while satisfaction partially mediated the effect on customer loyalty via trust. The findings can help managers of affluent banking services understand the important dimensions of customer personality in relational exchanges and develop relevant relationship management strategies to deliver satisfaction and enhance customer loyalty. In the recent years, the proliferation of financial technology, increased customer sophistication, and commoditisation of banking relationships have disrupted the banking industry. These days, retail banks face a challenging environment as consumers have a plethora of banking, wealth planning, and investment management options from licensed and restricted licensed banks , which explains the interaction between two parties during the exchange of resources Homans , has beeWithin an exchange relationship, feelings of trust can develop depending on the level of mutual reciprocation which happens when one party provides benefit to the other, and the other party returns the benefit at a later stage . Personality traits are archetypes where consumers express themselves and reflect the values, actions, and words of a consumer regarding a product, service, or firm , cognition  reported a positive impact of trust on satisfaction when examining the online shopping experience of retail customers. In recent studies, Rafiq et al.  and five items each  adopted from Anjam et al. . Trust wThe survey was pre-tested by professional bankers and academics with expertise in retail banking. Based on their suggestions, minor modifications were made. The revised survey was further pre-tested with four participants, namely two customers from the target group and two relationship managers from the local banks. The results of the pre-test confirmed the face validity and content validity of the instruments. Further, a pilot test was conducted with 52 graduate students. The pilot study established the reliability of the scales, as the values of Cronbach alpha, composite reliability (CR), and average variance extracted (AVE) were satisfactory, achieving construct reliability and validity , as all the items demonstrated loadings larger than 0.50 , cross-validated redundancy (Q2) and the path coefficients , 0.856 (satisfaction), and 0.813 (trust), which validated the strong predictive power of the model  of 0.04, presented a good fit between the conceptual model and the observed data. The predictive ability of the hypothesised model was examined using the following criteria: coefficient of determination  and satisfaction , were not supported. H2a, which proposed a positive link between conscientiousness and trust, was supported , but H2b, which proposed a positive correlation conscientiousness and satisfaction, was not supported . There was a positive link between extroversion and trust  and extroversion and satisfaction , providing support for both H3a and H3b. On the contrary, H4a and H4b, which suggested a positive connection between neuroticism and trust  and satisfaction  respectively, were both non-significant. H5a which proposed a positive association between agreeableness and trust  was supported, but H5b which suggested a positive link between agreeableness and satisfaction  was not supported. The association between trust and satisfaction was positive and significant , providing support for H6. Finally, H7, which posited a positive link between trust and loyalty  and H8, which proposed a connection between satisfaction and loyalty , were supported.The path coefficient and the As the conceptual model included the mediation effect of satisfaction between trust and loyalty, Baron and Kenny’s  mediatioF  = 13.78; p < 0.001, explaining 32% of the variance in customer loyalty. After the inclusion of trust and satisfaction in Block 2, the total variance increased to 88.80%, with F  = 176.03, p < 0.001. The introduction of trust and satisfaction explained an additional 57% of the variance in customer loyalty, after controlling for age, education level, occupation, gender, bank type, length of relationship, and frequency of contact (R2 Change = 0.57). The results showed that increased trust  and satisfaction  predicted greater customer loyalty. Among the demographic variables, length of relationship predicted customer loyalty as seen in the significant relationship between them .Hierarchical multiple regression analysis was performed to investigate the effects of trust and satisfaction on customer loyalty, after controlling for covariates such as age, education level, occupation, gender, bank type, length of relationship, and frequency of contact. Referring to Table The findings demonstrated the influence of the Big Five personality  and satisfaction (H1b) which contradict existing literature Castillo . As openSecond, both conscientiousness (H2a) and agreeableness (H5a) show positive relationships with trust which are consistent with previous studies Siddiqui . As consThird, the results verified the predictive power of extroversion on both trust (H3a) and satisfaction (H3b). This is not surprising, as extroverts are outgoing, sociable, active, chatty and prefer to engage with others, leading to perceptions of trust and satisfaction. In addition, an affirmative relationship between extroversion, trust, and satisfaction had been reported in previous studies Castillo , where hFourth, the findings did not establish any relationships between neuroticism and trust (H4a) nor satisfaction (H4b). Examining the research context, the relationships between neuroticism and trust/satisfaction were confirmed in non-banking contexts Castillo  but not Fifth, in terms of outcomes, trust displayed a significant influence on satisfaction (H6). This is supported by several studies which found trust and satisfaction as important determinants of customer loyalty  for extroverted and introverted customers. Branches offering a comfortable face-to-face banking experience which encourages social interactions could fulfil the needs of extroverts while virtual banks with full banking facilities could engage introverts.This study validated the effect of trust on both satisfaction and customer loyalty. Based on the importance of trust, managers should direct every effort to trust-building via professionalism which can promote long-lasting relationship in high contact, relational services (Balaji Rao and Rao In the era of Industry 4.0, banks can consider investing in AI and/or FinTech to help relationship managers in their actual service delivery Jaiwant . With thGiven the time and budget constraint, the study collected only 210 valid samples. Further studies can collect larger samples across premium banking services in different countries for better generalisation. The current study adopted a cross-sectional survey with a non-probability sampling method, thus future studies could consider longitudinal studies that can extend the scope of this study. This research focused solely on relational customers who have an existing relationship with their relationship manager. Future studies could examine whether transactional customers display similar relational attributes (Eriksson and Hermansson


H. jubatum under three low water potentials were measured. The results showed that the growth and photosynthetic parameters under these stresses were all decreased except for carotenoid (Car) with the increasing of stress concentration, and alkali stress caused the most damaging effects on the seedlings. The decreased net photosynthetic rate (Pn), stomatal conductance (Gs), and intercellular CO2 concentrations (Ci) values under salt stress were mainly attributed to stomatal factors, while non-stomatal factors were dominate under drought and alkali stresses. The reduced chlorophyll and slightly increased Car contents occurred under these stresses, and most significant changed under alkali stress. In addition, the maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ΦPSII), and photochemical quenching coefficient (qP) under the stresses were all decreased, indicating that salt, alkali, and drought stresses all increased susceptibility of PSII to photoinhibition, reduced the photosynthetic activity by the declined absorption of light for photochemistry, and increased PSII active reaction centers. Moreover, the non-photochemical quenching coefficient (NPQ) of alkali stress was different from salt and drought stresses, showing that the high pH of alkali stress caused more damaging effects on the photoprotection mechanism depending on the xanthophyll cycle. The above results suggest that the H. jubatum has stronger tolerance of salt than drought and alkali stresses, and the negative effects of alkali stress on the growth and photosynthetic performance of this species was most serious.Hordeum jubatum is a halophyte ornamental plant wildly distributed in the Northeast of China, where the low water potential induced by various abiotic stresses is a major factor limiting plant growth and development. However, little is known about the comparative effects of salt, alkali, and drought stresses at uniform water potential on the plants. In the present study, the growth, gas exchange parameters, photosynthetic pigments, and chlorophyll fluorescence in the seedlings of  The farmland physical and chemical soil properties were: 30.62% of soil water content, 16.61 g kg–1 of soil organic matter, 0.64 g kg–1 of total nitrogen, 0.05 g kg–1 of available nitrogen, 0.04 g kg–1 of available phosphorus, 0.11 g kg–1 of available potassium, and the pH is 7.02. The pots were cultivated in a greenhouse for germination, where the average temperature and light time were 25°C and 12 h, respectively.The mature seeds of 3) concentrations were also determined by Water Potential Meter , which were marked as A1, A2, and A3, respectively. In addition, three concentrations of PEG-6000 solution  were also configured to simulate drought stress based on the above water potentials  were applied, and the corresponding water potentials were −0.21, −0.47, and −0.82 MPa, respectively. To maintain uniform water potential, three alkali  was obtained. The water content (WC) of each treatment was calculated as the formula: (FW - DW)/FW × 100%.The fresh weight (FW) of shoot per pot was immediately weighed when seedlings of H. jubatum, four fully expanded leaves were randomly selected to measure gas exchange parameters per pot. The open flow LI-6400XT gas-exchange system  was used to determine the net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci), transpiration rate (Tr), and water use efficiency (WUE) between 9:00 and 11:30 am. The CO2 concentration, average temperature, flow rate and light flux intensity in the environment, and block temperature were 420 μmol mol–1, 25°C, 500 μmol s–1, 64 μmol m–2 s–1, and 28.5°C, respectively.After 46 days of seedling growth of After measuring various gas exchange parameters, nine leaves were randomly selected from each treatment to determinate chlorophyll fluorescence indicators using FluorCam FC 800-O fluorescence imaging system . Before measurement, all samples need to be dark-adapted for at least 30 min to ensure that the PSII reaction center is completely open. After dark adaptation, the samples were placed in FluorCam CCD camera, where four LED panels provided measuring flashes, actinic lights, and saturating pulses.P), and non-photochemical quenching coefficient (NPQ) were calculated according to The FluorCAM program was used to obtain a series of required fluorescence parameters by analyzing the fluorescence image of each treatment. The maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ΦPSII), photochemical quenching coefficient  at wavelengths of 663, 645, and 440 nm, respectively. The contents of chlorophyll (Chl a and Chl b) and carotenoid (Car) were calculated as the formulas described in H. jubatum seedlings, and the significant difference between treatments was analyzed by Duncan’s multiple range test (P < 0.05). Sigmaplot 13.0 software was used to plot the data between each treatment.The data used for comparison between each treatment are all the average values of each treatment, and SPSS 26.0  software was used for data comparison and analysis between each treatment. One-way analysis of variance (ANOVA) was used to analyze the average value and the comparative relationship between the treatments. Two-way ANOVA was used to analyze the effects of water potential, stress type and their interactions on the biomass and photosynthetic indexes of P < 0.01; Two-way ANOVA results showed that the FW, DW, and WC were significantly affected by water potential, stress type, and their interaction (Pn) and transpiration rate (Tr) were affected by water potential (P < 0.001), stress type (P < 0.001), and their interaction (P < 0.01). The stomatal conductance (Gs) and intercellular CO2 concentration (Ci) were significantly affected by water potential, stress type, and their interaction (P < 0.001). However, the WUE was only significantly affected by water potential . In addition, the Fv/Fm, ΦPSII were not affected by the interaction of water potential and stress type (P > 0.05), but the NPQ and qP were affected by the interaction of water potential and stress type . For example, the Fv/Fm of S3, A3, and D3 treatments decreased 13.0, 20.5, and 11.3%, respectively, compared to the control group (CK) at −0.82 MPa, and the ΦPSII of S3, A3, and D3 treatments decreased 31.7, 45.7, and 26.2%, respectively, under this water potential , but the qP of A2 and A3 treatments was more significantly declined than that of S2 and S3 treatments  and stress type (P < 0.01), while the Chl a was affected by the interaction of water potential and stress type (P < 0.05) and Chl b was not affected by their interaction (P > 0.05). Unlike Chl a and Chl b, the Car content was affected by water potential (P < 0.05) and stress type (P < 0.05), but not affected by the interaction of water potential and stress type  content and chlorophyll b (Chl b) content were affected by water potential  contents were all decreased under the three stresses, but the Car was slightly increased. The effect was the most obvious under the alkali stress . The mosP under different water potentials. In addition, the WUE and NPQ of salt stress had negative correlation with the growth parameters. For photosynthetic parameters, the Pn of salt stress had significantly positive correlation with Gs, Ci, Tr, and Chl b under different water potentials (P < 0.01). Moreover, the Fv/Fm of salt stress had significant positive correlation with ΦPSII, and the NPQ of salt stress had significant negative correlation with ΦPSII and Fv/Fm . Unlike salt stress, the WUE of alkali stress had no correlation with Gs and Tr under different water potentials  were all decreased under salt, alkali, and drought stresses with the decrease of water potential, indicating that photosynthesis of H. jubatum seedlings was significantly inhibited by these stresses  always estimates the redox status of the primary quinone electron acceptor of PSII (QA) and can also indicate the proportion of opened PSII reaction centers  on the donor side of PSII, and hindered the electron transfer on the PSII acceptor side (p values of the three stresses much decreased when the water potential was below −0.47 MPa, especially under alkali stress, indicating that the photosynthetic system was more easily influenced by stresses under lower water potential. Although the Fv/Fm, ΦPSII, and qp values under drought stress were slightly higher than those of salt stress, the differences were not significant, and the Pn value under salt stress was also significantly higher than that of drought stress, indicating that salt tolerance was stronger than drought tolerance of H. jubatum seedlings.Chlorophyll fluorescence can reflect the activity of the reaction center of the photosynthetic system by analyzing plant responses to light absorption and carbon assimilation processes . Fv/Fm, hibition . In the hibition . Thus, thibition . In addi centers . In thisi stress . This intor side . Moreove3 concentration disturbed the photoprotection mechanism of H. jubatum seedlings. It is concluded that the xanthophyll cycle depended on the proton gradient across the thylakoid membrane, the xanthophyll Zx and the PsbS protein plays a vital role in photoprotection of NPQ (Non-photochemical quenching can afford photoprotection to the photosynthetic apparatus by dissipating excess light energy that cannot be used for photochemistry in the form of heat under stresses . In the n of NPQ . TherefoH. jubatum occurred when subjected to these stresses. It hindered the photosynthesis process, especially under alkali stress. The reason may be that the high pH of alkali stress makes Fe, Mg, and other essential elements for chlorophyll synthesis reduced effectiveness (H. jubatum seedlings to dissipate excess light, resulting in a decrease of photosynthetic efficiency. Meanwhile, alkali stress also had the highest value in comparison to other stresses at the uniform water potential, indicating that the high pH of NaHCO3 caused more decline to the photochemical assimilation.The photosynthetic pigment content intuitively reflects the photosynthetic performance of plants under stress conditions . Chloroptiveness . MoreoveH. jubatum based on the same water potential. According to the photosynthetic gas exchange parameters, the decreased photosynthetic rate of H. jubatum under salt stress was mainly dominated by stomatal factors, but the non-stomatal factors played a major role under drought and alkali stresses. The chlorophyll content reduced under the three stresses reflected that the absorption and transmission of light energy in photosynthesis was degraded, among which was the most significant under alkali stress. Additionally, the decreased Fv/Fm, ΦPSII, and qP under these stresses indicated that salt, alkali, and drought stresses all increased susceptibility of PSII to photoinhibition, reduced the photosynthetic activity by the declined absorption of light for photochemistry, and increased PSII active reaction centers closed. Moreover, the NPQ was different from the salt and drought stresses under alkali stress, showing that the high pH caused by the NaHCO3 disturbed the photoprotection mechanism depending on the xanthophyll cycle. Overall, the H. jubatum had stronger tolerance of salt than that of drought and alkali, and the negative effects of alkali stress on growth and photosynthetic performance of this species were most serious.In summary, the result clearly analyzed the comparative effects of salt, drought, and alkali stresses on the growth, photosynthetic, and chlorophyll fluorescence performances in the seedlings of The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.CS wrote the manuscript and did the experiment. FY analyzed the data. ZL and XD did a part of the experiment. YL revised the grammar mistakes. JW and JL conceived and directed the manuscript. All authors contributed to manuscript revision and gave final approval for publication.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.


Pneumocystis jirovecii pneumonia (PcP) remains an important cause of morbimortality worldwide and a diagnostic challenge. Conventional methods have low accuracy, hardly discriminating colonization from infection, while some new high-cost or broncho-alveolar lavage-based methods have limited usefulness in developing countries. Quantitative PCR (qPCR) tests may overcome these limitations due to their high accuracy, possibility of automation, and decreasing cost. We evaluated an in-house qPCR targeting the fungus mtSSU gene using induced sputum. Sensitivity of the assay (ten target gene copies/assay) was determined using recombinant plasmids. We prospectively studied 86 AIDS patients with subacute respiratory symptoms in whom PcP was suspected. qPCR results were determined as quantification cycles (Cq) and compared with a qualitative PCR performed in the same IS, serum 1,3-β-D-Glucan assay, and a clinical/laboratory/radiology index for PcP. The qPCR clustered the patients in three groups: 32 with Cq ≤ 31 (qPCR+), 45 with Cq ≥ 33 (qPCR-), and nine with Cq between 31-33 (intermediary), which, combined with the other three analyses, enabled us to classify the groups as having PcP, not P. jirovecii-infected, and P. jirovecii-colonized, respectively. This molecular assay may contribute to improve PcP management, avoiding unnecessary treatments, and our knowledge of the natural history of this infection. Pneumocystis pneumonia (PcP) remains among the leading pulmonary opportunistic infections (OI) in several developing and developed countries [HIV is still an important cause of death across the world,  and Pneuountries ,4,5,6,7.ountries ,11,12,13ountries ,18,19,20ountries .P. jirovecii, low accuracy of the conventional diagnostic methods, and difficulties in sample collection. Methods based on visualization of the fungus with conventional staining have low sensitivity, requiring broncho-alveolar lavage fluid (BALF) samples for better performance. The commercially available immunofluorescence kits present higher sensitivity; however, they are not only generally expensive, but also require BALF samples for better results [PcP diagnosis remains a challenge, owing to several issues such as the absence of culture systems for  results ,25,26,27 results ,29.There are many reports on PCR assays for PcP diagnosis, with greater overall sensitivity. However, the use of different samples , in different settings , with different molecular techniques and gene target primers, have hampered the standardization and more widespread application of this technique ,39,40,411,3-β-D-glucan (BDG) dosage has also been applied in the diagnosis of PcP. However, due to its limitation of yielding non-specific results, the cutoff values for diagnosing PcP may be higher than usually recommended by the manufacturer, and the test may require confirmation through association with PCR assays ,58,59. FP. jirovecii in respiratory samples can indicate either PcP or colonization [This scenario is further complicated by the fact that AIDS patients often present concurrent pulmonary OIs, which lead to clinical and radiological features that overlap with those of PcP . In thesnization . OverallP. jirovecii in induced sputum (IS). We took into account the clinical characteristics of the patients, and compared our qPCR with the BDG assay and a conventional qualitative PCR assay (cPCR). IS has the advantage over “spontaneous” sputum because it retrieves more frequent lung samples from the upper bronchial tree and, compared with BALF sampling, is easier to perform and can be done at the bedside, since it does not require an invasive procedure [We thus carried out a prospective study in aids patients with suspected PcP of an in-house real-time PCR assay (qPCR) to quantitate rocedure ,63,64,653, and poor compliance to or not on ART and PcP prophylaxis. IS was collected before starting treatment for PcP (or with up to one dose), through inhalation of hypersaline solution (3% to 5% of NaCl), for 15–20 min, collected in a sterile container and stored at 4 °C until DNA extraction up to the next day. The study was approved by the IIER ethics committee (protocol 06/2016), and all patients signed an informed consent form.This was a prospective study that enrolled AIDS patients admitted between December 2017 and February 2020 at the emergency unit of the IIER, who were initially suspected of having PcP according to the following criteria: the presence of subacute cough and dyspnea (≥7 days), a current TCD4 count <250 cells/mm3; (d) LDH values greater than 350 UI/L; (e) O₂ saturation < 94.0% by pulse oximetry at room atmosphere; (f) positive clinical response to PcP treatment with cotrimoxazole or alternative regimens; (g) absence of treatment for other concurrent pulmonary diagnoses . In addition, the result of direct colorimetric examination performed in IS samples from a subset of patients was also considered: it was positive in eight patients out of the first 55 patients enrolled . Those with at least three of the above features were considered, based on our clinical expertise, as having enhanced likelihood for PcP , while those with less than three features were considered as having a low likelihood . This analysis was done blinded to the laboratory results by one of the authors (G.B.).In addition to the admission criteria, we defined specific features that enhanced the likelihood of PcP: (a) presence of dry cough (as opposed to productive cough); (b) interstitial infiltrates and/or ground-glass opacities on X-ray or CT-scan, respectively; (c) CD4 counts inferior to 100 cells/mmv/v in PBS pH 7.2, mixed, and shaken in a vortex. Then, tubes were centrifuged (2.500× g/10 min), and cell pellets were digested at 56 °C with 20 μL of proteinase K 20 mg/mL in 180 μL of AL buffer (Qiagen) until complete tissue digestion. The remaining steps were performed according to the manufacturer’s kit protocol. DNA samples concentration and purity were estimated by UV spectrophotometry  at 260 nm and 280 nm.One mL of the IS samples was used for DNA extractions, which were carried out with a QIAamp DNA mini kit . Sputum samples were previously dissolved P. jirovecii. PCR products (345 bp) were electrophoresed in 2% agarose gels, stained with ethidium bromide, and visualized under UV transilluminator apparatus  [The cPCR was previously established based on Khan et al., 1999 . Amplifi Israel) .P. jirovecii mitochondrial small subunit (mtSSU) gene, and resulting in 76 bp amplicon [The qPCR assay was performed with primers previously described, targeting the amplicon . A total6 to 1 × 100 copies/µL) in 20 independent experiments. The LOD was defined as the lowest number of copies detected in ≥ 95% of the experiments [The standard curves were constructed using recombinant plasmids obtained from the 76-bp PCR product cloned with the CloneJET PCR Cloning Kit , according to the manufacturer’s instructions. To establish the limit of detection (LOD), qPCR assays were performed with serial 10-fold dilutions of the recombinant plasmids  for BDG measurement according to the manufacturer’s instructions. The positivity cutoff was established at ≥ 150 pg/mL in our institution.Serum samples collected simultaneously to the IS were tested with the FungitellQualitative data were compared using Pearson’s Chi-square test and quantitative data was compared using ANOVA adjusted for white, and post-hoc analyses with Tukey’s test. Data analyses were performed using Integrated Development for R  . StatistThe standard curves showed linearity with a high correlation coefficient (R2 = 0.998) and an efficiency of 98%. The LOD of the qPCR assay achieved 10 copies of the recombinant plasmids/assay (detected in 100% of 20 independent experiments), corresponding to a mean Cq value of 32.01 ± 0.95 (CV = 2.98%), and a temperature melting of 76 °C ± 1 °C. The qPCR assay showed 100% analytical specificity, as no amplification signal was detected when all DNA samples from other fungal and bacterial strains were tested.A total of 97 PWH admitted to the emergency unit of the IIER with respiratory manifestations suggestive of PcP were enrolled. Of these, eight patients were excluded for being transferred to another health service in the first 24 h, and three for not being able to provide IS. Therefore, 86 patients underwent the radiology and laboratory evaluation prescribed by the attending physician. In addition, each patient provided on admission IS for DNA extraction and a blood sample for BDG quantification. Both were carried out later only for the study purpose: the results were not available to the attending physician. Both PCR assays were performed on the same DNA obtained from the IS samples as described in the P. jirovecii colonization only. It is of note that his/her Cq (31.0) was at the upper Cq limit of the qPCR-positive patients’ cluster. The third one  had a positive BDG test (310 pg/mL), but improved without specific treatment for PcP, which suggests colonization, although PcP could not be ruled out.Of the 86 patients enrolled in the study, the qPCR assay resulted positive (Cq < 33) in 41. We then analyzed the patients’ distribution according to their Cq values and high or low clinical index for PcP. P. jirovecii, since the BDG values in patients with positive (Cq ≤ 31) qPCR and high clinical index for PcP were almost invariably higher than 250 pg/mL. The other patient (#37) with a high clinical index for PcP also had a highly elevated BDG value (>500 pg/mL), and can therefore represent false-negative qPCR and cPCR results.On the other hand, among the 45 qPCR-negative patients, 33 were accordingly classified as unlikely PcP based on their low clinical index. The remaining 12 qPCR-negatives, with a high clinical index for PcP, were also diagnosed with a concurrent pulmonary infection  that could partly mimic the manifestations of PcP. They were all simultaneously treated for the concurrent pulmonary diseases and PcP, showing clinical improvement. However, 10 of these 12 patients had a negative BDG test, which, combined with the qPCR-negative  result, reinforced the unlikeliness of PcP. Of the two with a positive BDG test, one patient (#52), who had a BDG value (158 pg/mL) very close to our cutoff value, was treated for both PcP and CABP with clinical improvement. We considered the patient as not being infected with P. jirovecii colonization. The last patient (#100) had a low clinical index for PcP but a high BDG value (>500 pg/mL). However, his/her clinical follow-up also ruled out PcP: the patient was treated with levofloxacin alone, resulting in the symptoms’ resolution.Interestingly, of the nine patients in the intermediary qPCR group (31 > Cq < 33), seven had both low clinical index for PcP and a negative BDG test, which, combined with the high Ct values, led to the diagnosis of colonization. One patient had a high clinical index (#40) but undetectable BDG, and was treated for bacterial pneumonia alone, which resolved the respiratory manifestations, leading to the diagnosis of P. jirovecii: the patients #40 (Cq: 32.5) and #100 (Cq: 31.5) detailed above, and patient #50 (Cq: 32.7), with a low clinical index for PcP and negative BDG, resulting in the resolution of the respiratory symptoms without treatment for PcP. Thus, the nine patients with high Cq values (between 31 and 33) were probably colonized. Overall, the qPCR agreed with the cPCR, except for those patients with a positive qPCR with high Cq values.The cPCR was positive in all 32 positive (Cq ≤ 31) qPCR patients and negative in all 45 qPCR-negative patients, reinforcing the accuracy of molecular methods. In contrast, there was a pronounced disagreement in the intermediary group . This waWe also compared the qPCR with BDG results . The BDGIn the same way, of the 45 qPCR-negative patients, 40 were concurrently BDG-negative, ruling out PcP infection. Among the five with a discordant positive BDG value, there were two patients who were also detailed above: one of them (#52) with colonization, and the other (#37) with PcP and a probable false-negative qPCR result. The other three had positive BDG tests, but two patients (#42 and #93) had low clinical indices, were treated for an infection other than PcP pulmonary OI, and presented the resolution of the pulmonary symptoms, ruling out PcP. The remaining patient (#30) had a high clinical index, and was treated for disseminated TB and CABP, but died six days after admission; thus, as frequently happens in cases such as this, the diagnosis of PcP could neither be established nor ruled out.Finally, as mentioned earlier, among the nine patients in the intermediary group, eight were BDG-negative, and the single one (#100) with a positive BDG test was also diagnosed as colonization.p < 0.001). Of note, in the patients with a positive qPCR, there was a significant inverse correlation between the Cq and BDG values (p < 0.001 and r2 = −0.55).Overall, the BDG values were above the cutoff in 94% of the qPCR+ patients vs. 11% in both the qPCR- and colonization groups . A total of 90% of them were initially prescribed with anti-PcP treatment, a proportion significantly higher than the other groups. In fact, the qPCR+ group presented a median score of 5, significantly higher than the qPCR- (median: 2) and colonized groups (median: 1)  were qPCR+ with a Cq ≤ 31, 45 (52.3%) were qPCR-, and nine (10.5%) presented an intermediary Cq (between 31 and 33). Compared with the other two groups, the qPCR+ group displayed more frequently a set of clinical-laboratory parameters suggestive of PcP, which included dry cough, interstitial diffuse infiltrates on X-ray or “ground glass” on a CT scan, augmented DHL values, and decreased O2 saturation, together with greater immunosuppression (the lowest TCD4 count: median of 40 cells/mmdian: 1) . All of There are many reports on PCR assays for diagnosing PcP in patients with a variety of immunosuppressed conditions. Although they, in general, reported a good performance, they were not frequently included in the routine clinical practice. One of the most commonly reported limitations in these studies is the lack of a reliable gold standard method, such that the rate of true false-negative results is difficult to estimate . In addiP. jirovecii physiological state nor, in contrast to other genes, decreases with low fungal loads [P. jirovecii and PcP outbreaks have been reported [Another point that we believe contributed to the good performance of our qPCR assay was the choice of the target gene. We employed primers from the mtSSU ribosomal gene, which has been demonstrated to be present with a higher number of copies compared to the mtLSU gene. In addition, the number of mtSSU gene copies neither varies with al loads ,70. Thisal loads ,69,71,72al loads ,48,49,50al loads . In addireported ,73.P. jirovecii. All 45 qPCR- patients presented a concomitant negative cPCR, while the BGD test was negative in 40. Of the five patients with a positive BDG test, in three of them the presence of P. jirovecii was clinically ruled out, in one it was inconclusive, and in the other, the qPCR failed in diagnosing PcP.With regard to specificity, we found that our qPCR negative group, with one exception and a case whose diagnosis was inconclusive, was composed of patients not infected with The role of the BDG assay in the diagnosis of PcP has been intensely discussed: its major asset is its high negative predictive value, but its high sensitivity and lack of specificity due to factors that interfere with the tests ,75,76,77On the other hand, 12 of the 45 qPCR- patients presented a high clinical index for PcP. In all of these patients, pulmonary OIs other than PcP were diagnosed that could account for the respiratory manifestations, in agreement with the fact that concurrent pulmonary OIs are a rather common feature in AIDS patients. In total, nine of these patients had a negative BDG test, reassuring that PcP was not the cause of respiratory symptoms, and in one of them, the test was close to the cutoff value and considered by the clinical staff as not having PcP. The remaining two correspond to the false-negative (#37) and inconclusive diagnosis (#30) patients.The false-negative qPCR result in patient #37  points to an important but overlooked aspect of IS-based diagnostic methods that can lead to false-negative results. Our view is that the qPCR-negative result was possibly correct in that the fungus was truly absent from the IS sample, as we suspect was due to non-optimal IS collection. IS a simple procedure, but requires a standardized protocol, as well as the patient’s compliance, since hypertonic saline inhalation provokes mucosal irritation. The procedure yields specimens from the upper bronchial tree initially, and only after 15–20 min, samples from more peripheral airways . Proper The other issue is that the patients with a qPCR-positive test with higher Cq values, forming a cluster of results usually called the gray or intermediary zone. Our analyses of the nine patients who fell in this category showed that most of them had a low clinical index for PcP, a negative BDG test, and a negative cPCR in the same DNA sample, and were diagnosed as colonized patients. The two who did not strictly follow this rule had at least a low clinical index for PcP or a negative BDG test, but a clinical follow-up that ruled out PcP.P. jirovecii infections.Quantitative PCR assays are faster and easier to perform as an automated platform when compared with conventional PCR methods . We addiThis study has some limitations. Microscopic examinations were not available for most patients enrolled in our study due to the lack of specialized personnel. These methods usually require laboratory technicians with specific training. They are usually employed as the gold standard, especially when BALF samples are used; with IS, their sensitivity tends to decrease . As a reIn conclusion, this study shows that our in-house qPCR platform targeting the mtSSU gene from IS samples was able to discriminate three groups among AIDS patients according to the Cq values: one with PcP, one without PcP, and a third one with colonized patients. In this regard, this qPCR may represent an alternative method to microscopic examination in the diagnosis of PcP, as well as improve our knowledge of the natural history of this infection.


Purpose: Since the start of the coronavirus disease 2019 (COVID-19) pandemic in March 2020, ∼40% of U.S. adults have experienced delayed medical care. Rates of uninsurance, delayed care, and utilization of mental health services during the course of the pandemic have not been analyzed in detail. We examined monthly trends and disparities in access to care by household income levels in the United States.Methods: Using Census Bureau's nationally representative pooled 2020 Household Pulse Survey from April to December, 2020 , logistic regression models were used to analyze trends and inequalities in various access to care measures.Results: During the COVID-19 pandemic, the odds of being uninsured, having a delayed medical care due to pandemic, delayed care of something other than COVID-19, or delayed mental health care were, respectively, 5.54, 1.50, 1.85, and 2.18 times higher for adults with income <$25,000, compared to those with incomes ≥$200,000, controlling for age, sex, race/ethnicity, education, marital status, housing tenure, region of residence, and survey month. Income inequities in mental health care widened over the course of the pandemic, while the probability of delayed mental health care increased for all income groups. Although the odds of taking prescription medication for mental health were higher for low-income adults, the odds of receiving mental health services were generally lower for lower income adults, controlling for all covariates.Conclusion: In light of our findings on persistent health care inequities during the pandemic, increased policy efforts are needed to improve access to care in low-income populations as an equitable COVID-19 recovery response. Hamel et al., using a survey in June, 2020, found that individuals with income less than $40,000 were less likely to report delaying care due to coronavirus than those with income more than $40,000.19 The HPS utilized the Census Bureau's Master Address File as a source of sample, enabling to estimate at three different geographical levels, including Metropolitan Statistical Areas, state level, and national level.19 The HPS, as a short-turnaround instrument, provides employment status, spending patterns, food security, housing, physical and mental health, access to health care, and educational disruption among households in the United States during the COVID-19 pandemic.19 The HPS was initially developed on a weekly basis with three times of interviews for the same household in Phase 1, but Phase 2 and Phase 3 of the HPS consisted of 2-week cross-sectional datasets.19 For this study, to estimate monthly trend, we appended data of the weeks 1, 4, 7, and 10 from Phase 1, and weeks 13, 15, and 17 from Phase 2, and 19 and 21 from Phase 3, containing 9 months of HPS data during the pandemic from April to December, 2020. Human Participant Protection: The study was exempt from Institutional Review Board approval as it utilized a de-identified public use dataset.The data for this study are derived from the 2020 Household Pulse Survey (HPS), a nationally representative online survey developed by the U.S. Census Bureau with other federal agencies.The study sample comprised adults aged 18 to 88 in the 2020 HPS from April to December. The final sample size varied by the outcome measure ranging from 366,183 to 674,381  due to mSix outcome measures on access to care were used, including uninsured rate, delayed medical care due to pandemic, delayed care of something other than COVID-19, delayed mental health care, taking prescription medication for mental health, and receiving mental health services. The latter three outcome measures on access to mental health services were available from August to December. Respondents were asked (1) current health insurance coverage, and at any time in the last 4 weeks, whether they (2) “delay getting medical care because of the coronavirus pandemic?”; (3) “need medical care for something other than coronavirus, but did not get it because of the coronavirus pandemic?”; (4) “need counseling or therapy from a mental health professional, but did not get it for any reason?”; (5) “take prescription medication to help you with any emotions or with your concentration, behavior, or mental health?”; and (6) “receive counseling or therapy from a mental health professional such as a psychiatrist, psychologist, psychiatric nurse, or clinical social worker?.” All outcome measures were dichotomized with 1 being yes and zero equaling no.Household income was defined by six categories: less than $25,000; $25,000–$49,999; $50,000–$99,999; $100,000–$199,999; and $200,000 and above; missing. Although the original questionnaire has nine categories, we combined $25,000–$34,999, $35,000–$49,999 and $50,000–$74,999, $75,000–$99,999 to each category, given similar patterns in trend of access to care in the study sample.20–22 These covariates were measured as shown in Based on the previous literature and data availability, we selected the following covariates for model estimation: age, sex, race/ethnicity, education, marital status, housing tenure, region of residence, and survey month.19 The sample weights were adjusted by dividing by the number of pooling months. All analyses were conducted by Stata 16.23Logistic regression models were used to estimate the association between access to care and income level during the pandemic, controlling for individual characteristics, and month-fixed effects. Average marginal effects were calculated with delta-method standard errors. Complex survey design procedures were used to account for nonresponse, occupancy of the housing unit counts, the number of adults within the housing unit, and disproportionate sampling of demographic characteristics.p<0.05), except for the trend of uninsured rates for all income groups, and delayed mental health care and taking prescription medication for mental health for the highest income group (data not shown).Monthly trends of weighted unadjusted access measures for each group were statistically significant . The average predicted probabilities of delayed medical care due to the pandemic, delayed care of something other than COVID-19, and delayed mental health care were higher for adults with household income less than $25,000, compared to those with income $200,000 and above, by 9.1, 12.0, and 6.7 percentage points, respectively. The average predicted probability of taking prescription medication for mental health was higher for adults with household income less than $25,000, compared to those with income $200,000 and above by 10.2 percentage points. Although the average predicted probability of receiving mental health services was the highest for the lowest income group (12.52%), income groups between $25,000 and $199,999 presented a lower probability of receiving mental health services than those with income $200,000 and above (8.92–9.12% vs. 10.04%).2 or had no association with income level.3 Given that both reports used surveys from June 2020, and we found that the delayed medical care varied by month, differences in the findings might stem from the analysis of different time periods of the survey.Our study contributes to the existing literature by estimating the monthly trend of various access measures across income groups and by adding the evidence on the association between access to care and household income level among adults aged ≥18 years, using a nationally representative dataset from April to December during the COVID-19 pandemic in the United States. We found significantly higher adjusted odds of being uninsured, delayed care due to pandemic, delayed care of something other than COVID-19, and delayed mental health care among individuals with lower household incomes than those with higher household incomes. These results differ from findings of two recent reports that the delayed care during the pandemic was less likely among low-income adults25The odds of receiving mental health services were also higher for lower income groups than those for higher income groups, except the lowest income group . We found that the odds of taking prescription medication for mental health were higher for lower income groups than those for higher income groups. The higher odds of receiving mental health services among adults with household income less than $25,000 and taking prescription medication for mental health for lower income groups might be explained by their increased vulnerability to mental distress due to job insecurity and financial concerns during the pandemic.26 chronic pain,11 and acute cares7 or self-reported reasons of delayed care2 by income level or race/ethnicity. Recent studies found that cancer screenings, visits, therapy, and surgeries have significantly decreased and surgery for chronic pain were deferred during the pandamic.26 Given our findings, access to specialty care might also differ by household income level. Moreover, considering that utilization of health care services and health status were associated with neighborhood-level socioeconomic characteristics or racial/ethnic composition,27 supply of physician or specialty care medical professionals,28 or area deprivation,29 it would be worthwhile to examine the association between access to care during the pandemic with neighborhood-level characteristics.Further studies are needed to focus on delayed access to specialty care such as cancer,30–33 Policymakers should make an effort on removing barriers to the use of telehealth such as reimbursement or technology infrastructure.30 For example, the Centers for Medicare and Medicaid Services (CMS) extended the reimbursement for remote patient monitoring services for acute and chronic conditions during the public health emergency.31 However, the equitable access to telehealth should be established given socioeconomic and racial/ethnic inequalities in access to care.32 The CMS report found that beneficiaries with income more than $25,000 were more likely to have access to telemedicine appointments compared with those with income below $25,000.6 Clinical training and guidance for health care professionals are needed to increase skilled workforce who can switch delivery modes to telehealth.33To improve access to care during the COVID-19 pandemic, telehealth might be an alternative solution for individuals to obtain necessary medical care.15 Low-income individuals who reside in states, which opted out of the expansion of Medicaid eligibility, could have less access to medical care, compared with expansion states.35 Third, the timing of delayed care could be lagged since respondents were asked their experience in last 4 weeks preceding the survey. For example, week 1 data collected from April 23 to May 5 could include delayed care experienced in late March or early May. For consistency and comparability, we considered data collected during a month as the timing of delayed care in the trend graphs. Fourth, the respondents in HSP are more likely to be women and non-Hispanic whites and have a higher education, compared with the American Community Survey.24 Overrepresentation of non-Hispanic whites and those with higher education might result in an underestimate of the magnitude of income disparities in uninsured rates, delayed care, or health care utilization. We addressed disproportionate sampling of demographic characteristics by using survey weights, which rakes the demographics of the interviewed persons to education attainment/sex/age distributions and ethnicity/race/sex/age population distributions.19This study has limitations. First, since we used cross-sectional survey data, the estimated associations between income and access to care do not infer causality. However, since household income was measured as of 2019 but access to care in HPS was measured during the pandemic since March 2020, the estimates were less likely to be affected by reverse causality. Second, we did not control for state-level or county-level covariates, such as state's decision on Medicaid expansion, or county-level household income, unemployment rate, or physician supply.Using the nationally representative survey data from April to December 2020, we found a strong association between access to care and household income level among adults aged ≥18 years during the COVID-19 pandemic. Specifically, the likelihood of being uninsured rate and having delayed any medical care, care of something other than COVID-19, and mental health care was significantly higher for adults with lower household incomes than those with higher incomes. While the odds of taking prescription medication for mental health were higher for adults with low household incomes than those with high income, the odds of receiving mental health services were lower for lower income groups, except for the lowest income group, controlling for all covariates. During the pandemic, U.S. adults with lower household incomes had statistically significantly higher rates of uninsurance, delayed care, and the use of mental health services than those with higher income. These findings emphasize the need for increased policy efforts to improve access to care among low-income adults and households to reduce health care inequities during the ongoing pandemic and beyond.


Several recent studies revealed the significant contribution of intensive agriculture to global climate change and biodiversity decline. However, synthetic pesticides and fertilizers, which are among the main reasons for these negative effects, are required to achieve the high performance of elite crops needed to feed the growing world population. Modern agro-biologics, such as biopesticides, biostimulants, and biofertilizers are intended to replace or reduce the current agro-chemicals, but the former are often difficult to combine with the latter. Chitosans, produced from the fisheries’ byproduct chitin, are among the most promising agro-biologics, and copper fungicides are among the most widely used plant protectants in organic farming. However, the two active ingredients tend to form precipitates, hindering product development. Here, we show that partial hydrolysis of a chitosan polymer can yield a mixture of smaller polymers and oligomers that act synergistically in their antifungal activity. The low molecular weight (Mw) of this hydrolysate allows its combination with copper acetate, again leading to a synergistic effect. Combined, these synergies allow a 50% reduction in copper concentration, while maintaining the antifungal activity. This is potentially a significant step towards a more sustainable agriculture. Sufficient food production for a growing world population requires intensive agriculture, including effective plant protection measures. At the same time, the negative effects of the excessive use of chemical plant protectants on consumer health and the environment are increasingly apparent. To mitigate those by developing more sustainable agricultural practices, a reduction of chemical inputs is urgently required, but this needs to be accomplished without loss of efficacy. Despite their non-specific mode of action, copper-based fungicides are still in widespread use, pathogens do not develop resistance against them, and they are approved for use in organic agriculture. However, in the interest of a circular bioeconomy, the goal must be to reduce the copper-input in a field to an amount, which is later removed from the field with the harvest. To retain the antimicrobial efficacy at reduced dosage, copper fungicides may be combined with other active ingredients, ideally agro-biologics with no negative impact on the environment and for consumers. The agro-biologic with a potential for synergistic interactions with fungicides is chitosan. However, the combination of copper and chitosan is not trivial, as precipitates quickly form .N-acetyl-d-glucosamine (GlcNAc) units linked by β-1,4 glycosidic bonds, chitin is a frequent component of fungal cell walls and of the exo- or endoskeleton of many invertebrate animals. When some or all of the GlcNAc units in chitins are converted into d-glucosamine (GlcN) units via chemical or enzymatic de-N-acetylation, the resulting chitosans are polycationic molecules at slightly acidic pH values due to the protonation of the amino group. Unlike chitins, which form insoluble, crystalline fibers, chitosans are soluble at pH values below ca. 6, making them more attractive for a broad range of applications in diverse fields. Their non-toxicity towards animal and human cells, their biodegradability in the environment [As an aminopolysaccharide consisting of ironment , as wellironment ,4 and plironment ,6,7 woulironment ,9. As a ironment , but alsironment .FA) describes the proportion of acetylated GlcNAc units within the molecule, and thus determines its charge density. Most of the commercially available chitosans have FA values between 0.05 and 0.25. Together, DP and FA determine the Mw of the chitosan. The third, but by far less studied structural parameter, is the pattern of acetylation (PA), which describes the sequence of GlcNAc and GlcN units within the molecule. All of the commercially available chitosans appear to have random PAs. While the influence of DP and FA on antimicrobial and plant strengthening activities is now beginning to be understood, with chitosans of medium DP and low FA having the strongest antimicrobial activities [FA is best to induce plant disease resistance reactions [To date, both antimicrobial and plant resistance inducing activities of chitosans are known to be strongly dependent on the physicochemical properties of chitosans, which in turn depend on their chemical structure. Structurally, chitosan molecules are characterized by three parameters. First, the degree of polymerization (DP) describes the number of monomeric units in a chitosan chain, and thus determines its length. The boundary between chitosan oligomers and polymers is not clearly defined, but is typically assumed around DP 20. Second, the fraction of acetylation . However, the influence of this parameter on biological activities of chitosan samples has been disregarded until very recently. It was found that a chitosan sample with high Đ had higher antimicrobial activities than expected when adding the antimicrobial activities of its constituents, apparently due to a synergistic effect of the chitosan oligomers and polymers contained in the sample [However, similar to other polysaccharides and other polymers, chitosan samples are invariably mixtures of different chitosan molecules. Therefore, in addition to the three parameters described above, which define individual chitosan molecules, a chitosan solution is characterized also by its dispersity in DP, consists . The onle sample .Unfortunately, even the most efficient chitosan-based agro-biologics tend to perform well only under moderate inoculum pressure, when chitosan treatments often suffice to protect the crops from economically relevant losses, particularly in less intensive agricultural settings. However, in intensive Western-style agriculture, which requires optimal fertilization, water supply, and plant protection to realize the growth potential of high yielding elite varieties, the use of agro-biologics is often not sufficient, particularly in years with adverse climatic conditions. Currently, the potentially synergistic combination of agro-biologics with conventional agro-chemicals is emerging as a new strategy of integrated crop management, which promises to significantly reduce the input of chemicals in agriculture, even if not completely replacing them. This synergistic effect of structurally well-defined chitosans and synthetic fungicides has been well documented ,19, whilFA than in our earlier studies, as chitosan–copper interactions increase with the decreasing FA. Similar to our earlier studies [Fusarium graminearum, the causal agent of head blight in wheat, as a difficult to control phytopathogen of significant economic relevance [Our recent observation of a synergistic antimicrobial effect of small chitosan polymers and oligomers, which in combination exhibited even stronger effects than the most antimicrobially active polymers alone , may sug studies ,18, we uelevance . The ove1H–NMR spectroscopy and HP-SEC-RI-MALLS, we characterized the resulting chitosan (FA of 0.2 and a weight-average Mw of ca. 58.7 kDa (and thus a weight-average DP of ca. 350) with a Đ of 2.2 ), which is produced from shrimp shell chitin using a mild, one-step, semi-homogeneous alkaline deacetylation process . Using 1chitosan A as haviĐ of 2.2 .FA of 0.2 and a weight-average Mw of ca. 43.3 kDa (DP ca. 250) with a Đ of 1.1  ions was around 1 mM or ca. 60 µg mL−1, i.e., in a similar range as the chitosans  ion. The acetate salt was selected, as the chitosans were also solubilized using a 5% stoichiometric access of acetic acid. The IChitosans .50 value (1000 µM), (ii) a still antimicrobially active, but 50% lower concentration (500 µM), and (iii) an almost inactive, again 50% lower concentration (250 µM). These decreasing concentrations of copper were combined with increasing, but low, and thus non-to-barely active concentrations of chitosan hydrolysate , as we were aiming for a synergistic effect of the two active ingredients. To account for the slightly differing vigor of the fungus in different experiments, which leads to difficulties in comparing absolute inhibitory rates, the two ingredients were also included at their different concentrations alone.Next, we prepared three different mixtures of copper acetate with the chitosan hydrolysate, selecting concentrations based on the dose-response curves shown in F. graminearum compared to the earlier experiments. Growth inhibition by copper showed a clear dose-dependency, but the highest concentration used exhibited only about 40% inhibition. All of the three chitosan concentrations used were equally and only slightly inhibitory, while all of the three copper–chitosan combinations showed significantly stronger inhibitory activities than the respective copper or chitosan concentrations alone  combined with the lowest chitosan concentration used .In these experiments, both the copper(II) ions and the chitosan hydrolysate proved slightly less inhibitory against ns alone . The hig−1 chitosan hydrolysate was not significantly different from the positive control containing 2000 µM copper(II) ions and no chitosan. Similarly, the combination of 250 µM copper(II) ions with 80 µg mL−1 chitosan hydrolysate was as antimicrobially active as 1000 µM copper(II) ions alone. Clearly, the addition of low amounts of chitosan hydrolysate allowed a strong reduction in copper concentrations without loss of antimicrobial efficiency.Moreover, all of the three chitosan–copper combinations showed synergistic activity, with synergy factors ranging from 1.7 for the combination with the highest copper concentration to 2.5 for the combination with the lowest copper concentration . InteresF. graminearum, and that the antimicrobial activity is dependent on the DP and Đ of the chitosan solution used. Chitosan is known to reduce the growth of this pathogen both in vitro [in planta [Fusarium species causing diseases of potato plants and tubers by the application of chitosan [In this study, we verified that the chitosan treatment had direct antifungal effects on the vegetative growth of in vitro  and in vin vitro , and to n planta ,38. Recechitosan . A plethchitosan ,33. Our chitosan ,32. On tchitosan . Howeverchitosan . Most lichitosan .The mode of action of the antifungal effect of chitosans is not entirely understood. However, it is clearly dependent on both the target organism, e.g., its cell wall or plasma membrane composition or its chitosanolytic potential ,33,34, aFA 0.1, herein, we describe it for chitosans of FA 0.2, suggesting that it is a more general effect. The mechanism of this synergistic activity of chitosan mixtures is not yet understood. However, synergism is more likely to occur if the individual components of a mixture perform different modes of action on their targets. This tends to corroborate the above described mechanism of extracellular membrane disturbance by chitosan polymers, which leads to intracellular metabolic disturbance by chitosan oligomers. Even if we do not yet fully understand the mechanism of antifungal synergism of chitosan mixtures, the observation emphasizes the importance of considering the molecular weight dispersity (Đ) of a chitosan solution as a potentially crucial determinant of its biological activity [While the synergistic antimicrobial effect of chitosan polymers and oligomers was originally shown for chitosans of activity .FA is best [Given that extracellular chitosanolytic enzymes that are present in a target tissue might slowly process chitosan polymers into oligomers, this synergistic activity might even play a role in the efficacy of chitosan polymers alone. If we know the optimal structural features required for the bioactivity of the membrane-targeting chitosan polymers and those of the intracellularly acting chitosan oligomers, and if we know the sequence- dependency of the chitosanolytic enzymes that are present in a target tissue, it might be possible to design the chitosan polymer in a way that it disturbs membrane integrity, and, at the same time, acts as a source for the slow release of the bioactive oligomers. Membrane disturbance apparently requires high charge density, thus a very low  is best . However is best . Clearly is best .P. aeruginosa [Candida species [Phytophthora capsici [Botrytis cinerea [Neoscytalidium dimidiatum [Fusarium-caused tobacco root rot [FA 0.15) were combined with five different commercially available fungicides, resulting in synergistic antifungal activity against Botrytis cinerea, Alternaria brassicicola, and Mucor piriformis in both in vitro and in vivo studies, as well as in field trials [Beyond synergism between chitosan oligomers and polymers, we found an additional synergism of the hydrolysate and copper(II) ions. The antimicrobial synergistic activity of chitosan and a second compound—mostly a fungicide—have been described repeatedly. For example, mixtures of chitosan and essential oils have been reported to inhibit anthracnose diseases in papaya  and mangruginosa . Other sruginosa , as well species . Another capsici  and fung cinerea , Neoscytmidiatum , and Fusroot rot . In a syd trials . MoreoveCopper salts are among the oldest fungicides. However, they are still in frequent use due to their high efficacy and low chance of tolerance or resistance development in target organisms. The mode of action of copper-based fungicides is believed to be based on copper(II) ion uptake, followed by interaction with various chemical groups, most prominently sulfhydryl groups, ultimately disrupting the function of enzymes and other proteins , which eThis synergism between chitosan and copper has been previously reported. However, the formation of insoluble copper–chitosan complexes at higher pH values has long prevented the development of copper–chitosan products for plant protection, as copper exhibits phytotoxicity at the acidic pH value required for solubilization of the complexes. This problem can be overcome by nanoformulation, e.g., in the form of copper-loaded chitosan nanogels  or coppe2O and solubilization with a 5% molar excess of acetic acid relative to the free amino groups in the chitosan used. A partial acid hydrolysate of the chitosan was supplied by the same company as a 10% (w/v) solution. All of the solutions underwent sterile filtration through filters with a pore size of 0.22 μm before further usage.A chitosan polymer solution was obtained by dispersion of chitosan powder  in dHĐ = Mw/Mn was calculated, with Mw as the weight-average and Mn as the number-average molecular weight.Weight-average DP (DPw) of chitosan was determined using a combined system of high-pressure size exclusion chromatography coupled to refractive index detection and multi-angle laser light scattering analysis (HP-SEC-RID-MALLS), as described in . From thFA of chitosan, proton nuclear magnetic resonance spectroscopy (1H–NMR) was applied, according to a method described by [FA = (1/3 × CHI3)/(1/6 × I(H2 − H6)), with ICH3 as the integral of methyl group protons and I(H2 − H6) as the sum of integrals of H2, H3, H4, H5, and H6 protons of GlcN [To determine the ribed by . This me of GlcN .−1 solution of the chitosan hydrolysate was passed through a 0.45-µm filter and loaded into a loop with a capacity of 5 mL. The elution flow rate was maintained at 0.6 mL min−1. The effluent was monitored with an online refractive index detector  for around 27 h. After 8 h, fractions were collected every 10 min. After separating and collecting the desired fractions, the pooled fractions were lyophilized and re-solubilized in dH2O.Separation of polymer and oligomer fractions in the chitosan hydrolysate was conducted using semi-preparative size exclusion chromatography (semi-prepSEC) with a SECurity GPC system , along with a set of three HiLoadTM 26/60 SuperdexTM 30 preparatory grade columns (2.60 × 180 cm) equilibrated with filtered and degassed elution buffer  and a fraction collector . For separation, a 5 mg mLF. graminearum strain DSM 4528 was cultivated and proliferated in petri dishes containing complete medium (CM) [2O.Mycelium of ium (CM)  agar (Suium (CM)  (Supplem3 conidia per mL) or dH2O (blanks) was added to 150 μL CM, supplemented with 40 μL of a solution of chitosan, copper acetate (CuAc) or combinations of both compounds. The pH value of the medium was around 5.5. The plates were incubated under agitation as described above and fungal growth was recorded by UV–Vis spectrophotometric determination  of the optical density at 600 nm (OD600) every 24 h for a total of 96 h. In the case that a dose-dependent antifungal activity of different chitosans was compared, the half maximal inhibitory concentration (IC50) values were determined via GraphPad PRISM software . To achieve a sigmoidal curve, which is required for the calculation of IC50 values, artificial data points at very low and high concentrations had to be added, in some cases, to mark no or complete inhibition, respectively. These artificial data points are marked in grey in the respective figures if they fall into the concentration range of the x-axis shown. Statistical analysis was performed using one-way ANOVA and post-hoc Tukey test. All of the experiments were at least performed three times individually, with each experiment consisting of six technical replicates per agent.The antifungal activity of chitosan was measured in a 96-well microtiter plate, according to . AccordiCexp is the expected activity , which is calculated from the activities of the individual components A and B. If the ratio  of the experimentally observed activity (Cobs) and Cexp is greater than 1, the mixture shows synergistic activity, whereas a ratio close to 1 indicates an additive activity [To determine the potential synergy of chitosans and combinations of chitosan with CuAc, synergistic activity was calculated using Abbott’s formula, as follows:activity .In this study, we corroborated the importance of both the molecular weight and the molecular weight dispersity of a chitosan sample for its antifungal activity. Partial hydrolysis of a chitosan polymer can yield a synergistically acting mixture of polymer and oligomer molecules. This mixture can be combined with copper(II) ions, generating another synergistic effect. On a fundamental research level, these observations suggest different modes of actions of chitosan polymers and oligomers, and of chitosans and copper. On an applied development level, our study suggests a way for the reduction of copper dosages required for efficient plant protection without the need for nanoformulation, which supports the transition to a more sustainable agriculture.


These functional discrepancies of TG2 open form might be caused by its multifunctional nature, the existence of splicing variants, the cell type and stimulus, and the genetic backgrounds and variations of the mouse models used. TG2 is also involved in the phagocytosis of dead cells by macrophages and in fibrosis during tissue repair. Here, we summarize and discuss the multifunctional and controversial roles of TG2, focusing on cell death/survival and fibrosis.Transglutaminase 2 (TG2) is a ubiquitously expressed enzyme catalyzing the crosslinking between Gln and Lys residues and involved in various pathophysiological events. Besides this crosslinking activity, TG2 functions as a deamidase, GTPase, isopeptidase, adapter/scaffold, protein disulfide isomerase, and kinase. It also plays a role in the regulation of hypusination and serotonylation. Through these activities, TG2 is involved in cell growth, differentiation, cell death, inflammation, tissue repair, and fibrosis. Depending on the cell type and stimulus, TG2 changes its subcellular localization and biological activity, leading to cell death or survival. In normal unstressed cells, intracellular TG2 exhibits a GTP-bound closed conformation, exerting prosurvival functions. However, upon cell stimulation with Ca Water replaces the amine donor substrates, leading to the deamidation of glutamine. In addition, TG2 and factor XIIIa exhibit a Ca2+-dependent isopeptidase activity and can hydrolyze the isopeptide bond, at least under test tube conditions. TG2 exerts additional enzymatic activities that do not require Ca2+, i.e., it hydrolyzes ATP and GTP to mediate signal transduction through G-protein-coupled receptors, protein disulfide isomerases, and protein kinases, as well as interacts with several proteins as an adhesion or scaffold protein ). In addition, a free cytoplasmic GTP concentration higher than 100 μM is required to maintain the closed GTP-bound conformation of TG2, which inhibits its transamidase activity [A few studies have focused on the regulation of TG2 activity compared with the number of investigations on its transcriptional regulation. Caactivity .2+ and GTP are present at high and low levels, respectively. However, a highly oxidative state was reported to keep TG2 in the inactive state in the absence of stress due to the formation of disulfide bonds as a posttranslational modification [The conditions in the extracellular space are suitable for TG2 activation as Cafication , which wfication . Furtherfication ,81. TG2 fication . Finallyfication ,84.2+ requirement for TG2 activation, which might allow TG2 transamidase activity resulting from the conformational changes induced by locally increased Ca2+ levels [TGM2 is also involved in the regulation of the transamidase activity. Indeed, C-terminal-truncated variants lack part of or the entire GTP-binding pocket. Therefore, it is expected that the transamidase activity is not suppressed even by high GTP concentrations, resulting in an increased sensitivity and level of TG2 activation by Ca2+ under physiological conditions. The interaction between TG2 and membrane lipids might be another regulatory factor of TG2 transamidase activity. Sphingosylphosphocholine reduces the Ca+ levels . The altThe multifunctional activity, genetic variants, and conformational changes of TG2 complicate the understanding of its role in physiological and pathological events. The various functions of TG2, i.e., as a GTPase, PDI, kinase, and adapter/scaffold, and its role in transamidase activity are associated with both cell death and survival in various cellular environments. In addition, TG2 is localized in several subcellular spaces, such as the ECM, plasma membrane, cytosol, mitochondria, recycling endosomes, and nucleus, which influence its biological activities .TG2 accumulation has been demonstrated in various cellular and tissue types undergoing cell death . For exaPreviously, we reported that crosslinking of transcription factors by nuclear TG2 caused caspase-independent cell death. TG2 crosslinks and inactivates the general transcription factor Sp1, which results in a reduced expression of growth factor receptors, such as c-Met and EGF receptors, which are essential for cell survival ,92. AnotBy investigating liver diseases, we previously demonstrated that TG2 transamidase activity significantly increases in the nucleus of hepatocytes treated with alcohol or free fatty acid, promoting the crosslinking and inactivation of Sp1. The defect in Sp1 activity causes the downregulation of the hepatocyte growth factor c-Met, leading to caspase-independent hepatic cell death in cultured hepatocytes and animal models as well as in patients with alcoholic and non-alcoholic steatohepatitis ,91. ThisThe relationship between TG2 and cell death has been investigated in several neuronal models . TG2 expNε-(γ-glutamyl)lysine crosslinks and huntingtin aggregates in the nucleus [Transgenic mice overexpressing human TG2 selectively in neurons exhibited a dramatic increase in neuronal damage in the sensitive hippocampal regions after treatment with kainic acid, even though these mice presented no apparent phenotype in the absence of stress . Enhance nucleus , indicat nucleus ,123. Mut nucleus ,125,126, nucleus , dopamin nucleus ,125, pre nucleus , peroxis nucleus , and cyt nucleus . These r nucleus . It was  nucleus , in the  nucleus , and med nucleus .In addition to the regulation of cell death and survival, TG2 is involved in the activation of several immune cells such as dendritic cells , T cellsNε-(γ-glutamyl)lysine isopeptide bond resulting from the crosslinking. This is an important step for the maturation and stabilization of ECM components, such as collagens, exacerbating scarring and fibrosis in various tissues, including the liver [2+-dependent early entrenchment  by crosslinking of the extant matrix, whereas LOX implicates Ca2+-independent contractility at later times [The TG2 transamidase activity contributes to the wound healing process and fibrosis. The reaction products form an he liver ,152,153,he liver ,158,159,he liver ,164,165,he liver ,167. Thehe liver . Impairehe liver . In the er times . These rAside from the direct ECM stabilization, the TG2 transamidase activity appears to play a significant role in the fixation and activation of the profibrotic cytokine TGF-β. TGF-β is released in a latent form and converted to an active one. Enhanced TG2 activity is required for TGF-β activation from the latency binding complex as it promotes crosslinking of the large latent TGF-β binding protein to fibronectin or other ECM components on the cell surface ,173,174.As previously described, we demonstrated that nuclear TG2 inactivated Sp1 by crosslinking, leading to reduced expression of c-Met and consequently activation of hepatic apoptosis in a hepatic injury mouse model and in patients with alcoholic steatohepatitis . TG2-medTGM2 gene, in the mouse background, and in the disease model. We obtained similar results using TG2-deficient mice. Indeed, liver fibrosis induced by bile duct ligation was not inhibited in these mice [However, TG2-deficient mice demonstrated no alteration of the fibrosis levels in the liver after treatment with CCl4 or thioacetamide . This coese mice , althougese mice  and lungese mice . Interesese mice ,188,189.2+ levels and high GTP concentrations in normal cellular condition without stress, TG2 exists as a closed form and exerts GTPase, PDI, and kinase activities to maintain homeostasis, which is crucial for cell survival. Contrarily, abnormal cellular conditions with stress and high Ca2+ concentrations allow TG2 change of conformation to the open form, which exhibits crosslinking activity, leading to alterations of its subcellular localization and extracellular release. Activated TG2 in the cytoplasm or nucleus of stressed cells crosslinks and regulates the activity and proteostasis by ubiquitination and autophagy of a number of target substrate proteins involved in several signaling events. Nuclear TG2 mainly regulates the activity of transcription factors and chromatin remodeling, which are involved in the expression of various downstream proteins important in cell death and survival.Since the TGase family is multifunctional and contains numerous isozymes and splicing variants, an integrated understanding of TGase functions in pathophysiological events is often difficult to achieve. The role of TG2 in cell death and survival remains controversial. However, TG2 may be generally involved in the positive effect of apoptotic cell phagocytosis by macrophages and fibrosis induction. At low CaIn addition to the activation of TG2 during cell death induction, TG2 is upregulated in macrophages and is important for the clearance by phagocytosis of dead cells. After or following cell death and inflammation events, TG2 is secreted into the extracellular space and contributes to the enhancement of fibrogenesis, which allows filling the gaps resulting from cell death. In the ECM, TG2 forms a complex with fibronectin and interacts with integrins and heparan sulfate proteoglycans, contributing to the stabilization of the ECM mediated by crosslinking and to sustained TGF-β activation, leading to the development of fibrosis linked to organ dysfunctions .Treatment with inhibitors of the TG2 crosslinking activity appeared to consistently suppress pathogenic fibrosis, although their effects on cell death and survival are not consistent, probably due to different experimental conditions and stimuli. Considering the ubiquitous and multifunctional nature of TG2, the development of a drug with no side effects might be difficult. Therefore, clinical compounds and antibodies that are more specific and allow controlling the drug distribution in the whole body are currently being developed. Effective TG2 inhibitors were developed by Zedira GmbH and are now in advanced clinical trials for the treatment of celiac disease. Recently, novel candidates for the treatment of kidney fibrosis were also successfully developed. In the future, these TG2 inhibitors will also be tested in patients with fibrosis in other organs such as lung and liver.In our recent work, we conducted a comprehensive analysis of TG2-mediated crosslinking of substrate proteins in models of fibrosis targeting the liver, kidney, and lungs. Based on these results, we will create a database of the substrate proteins crosslinked in fibrosis, specifically, in each organ, which will support the development of novel preventive drugs against fibrosis acting by suppressing the crosslinking of substrate proteins.


Ti-based alloys are an important class of materials suitable especially for medical applications, but they are also used in the industrial sector. Due to their low tribological properties it is necessary to find optimal technologies and alloying elements in order to develop new alloys with improved properties. In this paper, a study on the influence of sintering treatments on the final properties of a titanium alloy is presented. The alloy of interest was obtained using the powders in following weight ratio: 80% wt Ti, 8% wt Mn, 3% wt Sn, 6% wt Aluminix123, 2% wt Zr and 1% wt graphite. Two sintering methods were used, namely two-step sintering (TSS) and multiple-step sintering (MSS), as alternatives to conventional sintering which uses a single sintering dwell time. Evolution of sample morphology, composition and crystalline structure with sintering method was evidenced. The lower values for the friction coefficient and for the wear rate was attained in the case of the sample obtained by TSS. Titanium has some spectacular properties, such as high toughness, low density and good corrosion resistance, making it attractive to be used most frequently in the biomedical field ,2,3. How2 powders can improve sintered density, induce microstructure modification, and reduce the oxygen content and cost of raw materials, compared with Ti powders [2, TiH2) [Some researchers have found that the use of TiH powders . Therefo2, TiH2) .However, the high friction coefficient and lower wear resistance than steel and Ni -based alloys have been a major drawback to its practical applications ,22. In tZr has chemical properties similar to Ti, and the formation of solid solutions causes the hardening of the alloy, increases the corrosion resistance and improves biocompatibility ,28. AlthAccording to Lu et al.  the addi3) and combination of moderate strength and high ductility, thermal conductivity, electric conductivity, corrosion-resistance [The addition of aluminum in Ni-Ti alloy researched by Pavel Salvetr et al. showed the decrease of porosity and improvement of corrosion resistance and hardness . Aluminusistance ,35.Previous studies concerning sintering procedure have shown that two-step (TSS) and multiple-step (MSS) sintering techniques allow researchers to obtain homogeneous compositions and microstructures of Ti-based alloys at low cost . The proThe selection of the processing methods is a critical factor that influences the microstructure, phase formation and its kinetics, as well as mechanical and tribological properties of any material. Therefore, the final properties and suitability to different applications can be tailored by fine-tuning of a selected production technique . PreviouTherefore, in order to optimize the sintering process of the titanium-based alloy, it was considered of interest to use the sintering temperature of 1050 °C, with the increase of the dwell time to 100 min. At the same time, in order to find more economical solutions in terms of energy and the total cost of obtaining the titanium-based alloy, the two types of treatments presented in this paper were chosen: TSS and MSS.The stated goal of this research is to find cheaper technologies for obtaining titanium-based alloy, in order to eliminate the main obstacle in the use of titanium, especially in the automotive field, namely the high cost of pure titanium. Therefore, it acted in two directions: (i) the use of titanium hydride as a raw material and (ii) the use of cheaper but more modern technologies, compared to classical sintering.2, 8% wt. Mn, 3% wt. Sn, 6% wt. Alumix321, 2% wt. Zr and 1% wt. graphite. This Ti-based alloy was obtained by PM and was subjected to two types of sintering processes: TSS and MSS. The main purpose of this work was to identify the most appropriate sintering procedure which allows the decrease of the manufacturing cost and the optimal properties for industrial applications, respectively, for components in the automotive industry.Given the previous results obtained by adding small amounts of metallic elements that allowed to obtain Ti-based alloys with improved final properties, we developed a Ti-based alloy with the following chemical composition: 80% wt. TiH-2) with particle size min. 99.9% <63 µm, water atomized produced by Chemetall GmbH , as a basic metal matrix, having titanium min. 95%, hydrogen min. 3.8%, small amounts of Si, Mg, Ni, Al, Fe max. 0.1% for each of them;Micronic hydride powder  provenience, purity 99.5% used for improving the wear behavior in friction conditions due to its property to form a protective layer of oxide in contact with air ;-Tin micronic powder, SN006020, produced by Goodfellow company ; tin helps titanium in the grinding process and is a metal with low influence in the transformation temperature; it also behaves very well in friction conditions ;-Zirconium nanometric powder, purchased from the Goodfellow company . Zirconium has the same atomic structure as titanium, being considered an excellent alloying element of titanium, with a good corrosion resistance and has influence on reducing the transformation temperature ,42;-Graphite powder, 2N5 type from American Elements company , having 99.5% purity and low friction coefficient. Due to Van der Waals forces, graphite has a very good wear resistance, being an excellent dry lubricant, these being the reasons why it was added in the composition of the alloy ;-Alumix powder Al 123, based on aluminum powder but with small amounts of 4.5% wt. Cu, 1.31% wt. Mg, 0.5% wt. Si, 0.10% Fe, 0.05% wt. Sn 2%.The following raw materials were used in the production procedure:2, 8%wt. Mn, 3%wt. Sn, 6%wt. Alumix321, 2%wt. Zr and 1%wt. graphite.The selected powders were blended and mechanically alloyed under Ar atmosphere for 120 min using a planetary mono mill PULVERISETTE 6 from Fritsch,  . The weight ratio between the components were: 80%wt. TiHIn order to obtain the green parts (not sintered), two grams of mixture were weighted using a Partner WPS 510/C/2  balance, followed by the uniaxial pressing process with a 600 MPa pressure, at room temperature and without lubrication. For the compaction process, a X200Cr120 cylindrical die was used.For the green compact’s densification, two sintering treatments were chosen, as shown in -sintering in two steps (TSS), with heating up to 1050 °C as the first stage, dwell time of 25 min and then the second sintering stage at 950 °C, with a dwell time of 75 min.-sintering in multiple steps (MSS), the first sintering level being at 1050 °C, with a dwell time of 25 min, the second sintering level at 1000 °C, with a dwell time of 20 min and the third sintering level at 950 °C, with a dwell time of 55 min.According to The heating rate was 6 °C/min for both sintering types. The green compacts were heated into a Nabertherm L3/11/C6  furnace in Ar atmosphere.After sintering, the samples were cooled down to room temperature in the furnace.The morphology and elemental analysis of the precursor powders and sinters were evidenced using a scanning electron microscope (SEM) from Hitachi  coupled with energy dispersive X-ray spectroscopy (EDXS) . For this purpose, the TSS and MSS specimens were axial sectioned using a Metkon Metacut-M 250  cut-off machine and heat-mounted in transparent resin, using a Metkon Metapress-A  mounting press. After that, the samples were grinded with silicon carbide papers with the following grits: 80, 120, 240, 400, 800, 1200 and 2000. Finally, the samples were polished with a GALAXY Polishing cloth PHI  and Dia-Duo 2 suspension . For grinding and polishing processes, a Metkon Forcipol 2V  machine was used.In order to study the particle size distribution of powders, the dynamic laser scattering (DLS) technique was used, and the equipment was a Brookhaven 90Plus by Brookhaven Instruments Corporation . The samples were dispersed in water and subjected to the ultrasonication for 5 min.X-ray diffraction (XRD) was used to perform the crystalline phase analysis in the sintered bodies .Tribological tests were performed with a TRB 01-2541 tribometer, using TriboX software , while a Surtronic S25 Taylor Hobson Profilometer  was used to measure the worn track section. In these cases, a steel ball 440C was used as counterpart and titanium-based alloy samples with a diameter of 12 mm obtained by TSS and MSS were used as tested parts. The testing conditions were: testing type—linear, normal load—2N, amplitude—6 mm, testing distance—24 m, atmosphere—air.After the friction coefficient measurements, the worn track sections of the samples and the balls used were analyzed using a Nikon MA 100 microscope  equipped with NIS ELEMENTS software .loss is the volumetric loss of the sample, F is the force applied and D is the total sliding distance.In order to calculate the wear rates, the worn track sections were measured using the Surtronic 25+ profilometer , and the formula used is given by:2 and Mn powders , is shown in In 2 precursor powder has suffered a dehydrogenation process leading to Ti-based alloy. After sintering, the variations in sample height and diameter during shrinkage were measured. In During the sintering process, the TiHAfter compaction and sintering, the green and the final density of the parts have been established by measuring their dimensional sizes and weighing them with an analytical balance, and then, the density formula for cylindrical parts has been applied. In In 2 dehydrogenation during the sintering treatment is indicated by the presence of pure titanium grains in high ratio .Using XRD analysis, crystalline phases present in the investigated samples were evidenced. Thus, in the sample obtained by TSS a three cIn 3) than the density of the MSS sample (3.61 g/cm3).As −3 µm, and for the sample obtained by MSS, the value of the penetration depth is −9.607 × 10−3 µm.The average of the penetration depth in the case of the sample obtained by the TSS method is −9.596 × 10In In From The samples obtained by the TSS process present better wear behavior in comparison with the ones obtained by MSS process due to the highest values of the densities of TSS samples leading to better mechanical properties. The average value of the wear coefficients is similar and fit within the values indicated in other studies dedicated to titanium-based alloy ,48.A titanium-based alloy with superior wear behaviour has been obtained using two sintering procedures. The structural analysis of the resulted sinters shows a relatively homogeneous distribution of the alloying elements, as well as the simultaneous presence of several crystalline phases. Two-step sintering or multiple-step sintering ensures lower energy consumption, which is a great advantage from an economic point of view compared to conventional sintering. The resulting alloy has a higher wear behavior than most titanium alloys. Friction coefficient values of 0.545 for the TSS samples, and 0.567 for those obtained by MSS, indicate a material with good antifriction properties. Due to its low weight, compared to the most used materials in the automotive area and its resistance to wear, the applicability of the as-obtained Ti-based alloy was configured in the field of automotive components, particularly the manufacturing of a connecting rod.This research opens future works which will be focused on the decreasing of the particle size and to study the influence of different sintering treatments, such as spark plasma sintering (SPS) and microwave sintering (MWS) on the properties of Ti-based alloy. Also, Ti-based alloy as ultrafine powders can be used in the additive manufacturing (AM) process, in order to produce parts for automotive industry.