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Diagnostics ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 196
Author(s):  
Junji Hosokawa-Muto ◽  
Yukiko Sassa-O’Brien ◽  
Yoshihito Fujinami ◽  
Hiroaki Nakahara

When examining infectious samples, rapid identification of the pathogenic agent is required for diagnosis and treatment or for investigating the cause of death. In our previous study, we applied exhaustive amplification using non-specific primers (the rapid determination system of viral genome sequences, the RDV method) to identify the causative virus via swab samples from a cat with a suspected viral infection. The purpose of the current study is to investigate suitable methods for the rapid identification of causative pathogens from infected tissue samples. First, the influenza virus was inoculated into mice to prepare infected tissue samples. RNA extracted from the mouse lung homogenates was transcribed into cDNA and then analyzed using the RDV method and next-generation sequencing, using MiSeq and MinION sequencers. The RDV method was unable to detect the influenza virus in the infected tissue samples. However, influenza virus reads were detected using next-generation sequencing. Comparing MiSeq and MinION, the time required for library and sequence preparation was shorter for MinION sequencing than for MiSeq sequencing. We conclude that when a causative virus needs to be rapidly identified from an infectious sample, MinION sequencing is currently the method of choice.


2021 ◽  
Vol 12 ◽  
Author(s):  
Zhongxia Tan ◽  
Youxi Zhang ◽  
Chao Wang ◽  
Le Sun

The aim of this study was to develop physiologically based pharmacokinetic (PBPK) models capable of simulating cefadroxil concentrations in plasma and tissues in mouse, rat, and human. PBPK models in this study consisted of 14 tissues and 2 blood compartments. They were established using measured tissue to plasma partition coefficient (Kp) in mouse and rat, absolute expression levels of hPEPT1 along the entire length of the human intestine, and the transporter kinetic parameters. The PBPK models also assumed that all the tissues were well-stirred compartments with perfusion rate limitations, and the ratio of the concentration in tissue to the unbound concentration in plasma is identical across species. These PBPK models were validated strictly by a series of observed plasma concentration–time profile data. The average fold error (AFE) and absolute average fold error (AAFE) values were all less than 2. The models’ rationality and accuracy were further demonstrated by the almost consistent Vss calculated by the PBPK model and noncompartmental method, as well as the good allometric scaling relationship of Vss and CL. The model suggests that hPEPT1 is the major transporter responsible for the oral absorption of cefadroxil in human, and the plasma concentration–time profiles of cefadroxil were not sensitive to dissolution rate faster than T85% = 2 h. The cefadroxil PBPK model in human is reliable and can be used to predict concentration–time profile at infected tissue. It may be useful for dose selection and informative decision-making during clinical trials and dosage form design of cefadroxil and provide a reference for the PBPK model establishment of hPEPT1 substrate.


2021 ◽  
Author(s):  
Holly C. Webster ◽  
Virginia Gamino ◽  
Amy L. Shergold ◽  
Anna T. Andrusaite ◽  
Graham A. Heieis ◽  
...  

Type 2 immunity is activated in response to both allergens and helminth infection. It can be detrimental or beneficial, and there is a pressing need to better understand its regulation. The immunosuppressive cytokine IL-10 is known as a T helper 2 (Th2) effector molecule, but it is currently unclear whether IL-10 dampens or promotes Th2 differentiation during infection. Here we show that helminth infection in mice elicits IL-10 expression in both the intestinal lamina propria and the draining mesenteric lymph node, with higher expression in the infected tissue. In vitro, exogenous IL-10 enhanced Th2 differentiation in isolated CD4+ T cells, increasing expression of GATA3 and production of IL-5 and IL-13. The ability of IL-10 to amplify the Th2 response coincided with its suppression of IFNγ expression and, in vivo, we found that, in intestinal helminth infection, IL-10 receptor expression was higher on Th1 cells in the small intestine than on Th2 cells in the same tissue, or on any Th cell in the draining lymph node. In vivo blockade of IL-10 signalling during helminth infection resulted in an expansion of IFNγ+ and Tbet+ Th1 cells in the small intestine and caused a coincident decrease in IL-13, IL-5 and GATA3 expression by intestinal T cells. Together our data indicate that IL-10 signalling promotes Th2 differentiation during helminth infection at least in part by regulating competing Th1 cells in the infected tissue.


2021 ◽  
Vol 06 (02) ◽  
pp. e63-e69
Author(s):  
Haruo Ogawa ◽  
Haruki Nakayama ◽  
Shinichi Nakayama ◽  
Shinya Tahara

Abstract Background Necrotizing fasciitis is a well-known disease that causes extensive tissue infection and requires radical debridement of the infected tissue. It can occur in all parts of the body, but there are few reports of necrotizing fasciitis in the axilla. We treated three patients with axillary necrotizing fasciitis. Methods In all cases, patients were referred to us after radical debridement of the infected soft tissue in the emergency department. At the first visit to our department, there were fist-sized soft tissue defects in the axilla. Moreover, the ipsilateral pectoralis major and latissimus dorsi muscles were partially resected because of the debridement of necrotizing fasciitis. In all cases, the ipsilateral thoracodorsal vessels were severely damaged and free-flap transfer was performed to close the axillary wound. Results All free flaps survived without complications. The patient's range of motion for shoulder abduction on the affected side was maintained postoperatively. Conclusion If necrotizing fasciitis occurs in the axilla, tissue infection can spread beyond it. In such a case, free-flap transfer can be an optimal treatment. Radical resection of the infected tissue results in the absence of recipient vessels in the axilla. Surgeons should bear in mind that, because of radical resection of the infected tissue, they may need to seek recipient vessels for free-flap transfer far from the axilla.


2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Leonardo Mancini ◽  
Alessia Pisaneschi ◽  
Vincenzo Mancini ◽  
Marino Ginoble ◽  
Vincenzo Quinzi ◽  
...  

Tooth decay is a multifactorial disease. Fermentable sugar, host factors, and cariogenic microbial flora are several agents that influence dental structure. In literature, alternative protocols for treating cavities are always of interest in terms of reducing pain and preserving tissue. In this case report, the use of a new gel-based on papain, which has a chemical effect on bacteria and allows the consistency of the altered tissue to be modified, leads to a less traumatic removal of the infected tissue. In this case report, BRIX3000, a gel with papain as its main ingredient, was used to treat an interproximal cavity on an upper premolar of a 35-year-old man frightened of the turbine. After a clinical check of all the systematic conditions and a first view of the oral cavity, the protocol was explained to the patient, and informed consent was obtained. The protocol involved applying the papain gel directly to the cavity, and after 2.5–3 minutes, it was removed. The complete removal of the infected tissue was achieved in three steps. The patient reported no discomfort, and the cavity was completely treated and ready to be restored. In conclusion, BRIX3000 seems to be a good alternative to the turbine in cavity treatment, particularly in patients who have discomfort during traditional treatments and are frightened of the turbine.


2021 ◽  
Author(s):  
Shannon Catherine Duffy ◽  
Manigandan Venkatesan ◽  
Shubhada Chothe ◽  
Indira Poojary ◽  
Valsan Philip Verghese ◽  
...  

Mycobacterium bovis bacille Calmette-Guérin (BCG) is a live attenuated vaccine which can result in local or disseminated infection, most commonly in immunocompromised individuals. Differentiation of BCG from other members of the Mycobacterium tuberculosis complex (MTBC) is required to diagnose BCG disease, which requires specific management. Current methods for BCG diagnosis are based on mycobacterial culture and conventional PCR; the former is time-consuming and the latter often unavailable. Further, there are reports that certain BCG strains may be associated with a higher rate of adverse events. This study describes the development of a two-step multiplex real-time PCR assay which uses single nucleotide polymorphisms to detect BCG and identify early or late BCG strains. The assay has a limit of detection of 1 pg BCG boiled lysate DNA and was shown to detect BCG in both pure cultures and experimentally infected tissue. Performance was assessed on 19 suspected BCG clinical isolates at Christian Medical College in Vellore, India taken from January 2018 to August 2020. Of these 19 isolates, 10 were identified as BCG (6 early and 4 late strains) and 9 were identified as other MTBC members. Taken together, the results demonstrate the ability of this assay to identify and characterize BCG disease from cultures and infected tissue. The capacity to identify BCG may improve patient management and the ability to discriminate between BCG strains may enable BCG vaccine pharmacovigilance.


2021 ◽  
Vol 15 (4) ◽  
pp. e0009295
Author(s):  
Rogger P. Carmen-Orozco ◽  
Danitza G. Dávila-Villacorta ◽  
Ana D. Delgado-Kamiche ◽  
Rensson H. Celiz ◽  
Grace Trompeter ◽  
...  

Background The parasite Taenia solium causes neurocysticercosis (NCC) in humans and is a common cause of adult-onset epilepsy in the developing world. Hippocampal atrophy, which occurs far from the cyst, is an emerging new complication of NCC. Evaluation of molecular pathways in brain regions close to and distant from the cyst could offer insight into this pathology. Methods Rats were inoculated intracranially with T. solium oncospheres. After 4 months, RNA was extracted from brain tissue samples in rats with NCC and uninfected controls, and cDNA was generated. Expression of 38 genes related to different molecular pathways involved in the inflammatory response and healing was assessed by RT-PCR array. Results Inflammatory cytokines IFN-γ, TNF-α, and IL-1, together with TGF-β and ARG-1, were overexpressed in tissue close to the parasite compared to non-infected tissue. Genes for IL-1A, CSF-1, FN-1, COL-3A1, and MMP-2 were overexpressed in contralateral tissue compared to non-infected tissue. Conclusions The viable cysticerci in the rat model for NCC is characterized by increased expression of genes associated with a proinflammatory response and fibrosis-related proteins, which may mediate the chronic state of infection. These pathways appear to influence regions far from the cyst, which may explain the emerging association between NCC and hippocampal atrophy.


2021 ◽  
Author(s):  
Sebastjan Radišek

Abstract Hop stunt viroid (HSVd) is a covalently closed, single-stranded RNA molecule of 297 nucleotides (Sano et al., 1985). Variants consisting of 294-303 nucleotides have been described from different hosts (Kofalvi et al., 1997; Amari et al., 2001). HSVd has a rod-like conformation with five domains, a central conserved region (CCR) similar to members of genera Pospiviroid and Cocadviroid, and a terminal conserved hairpin (TCH) which is also present in the genus Cocadviroid (Flores et al., 1997). Replication is known to occur through an asymmetric, rolling-circle model because longer-than-unit minus strands have been found in infected tissue (Flores et al., 2005). HSVd was named due to the first identification of the pathogen on hop plants originating from Japan (Yamamoto et al., 1973). In a wide range of host species, infection by HSVd appears to be latent, whereas in hop, Citrus and Prunus species it causes specific disorders and economic damage (Hadidi et al., 2003).


Breast Care ◽  
2021 ◽  
pp. 1-8
Author(s):  
Huiying Xu ◽  
Ruidong Liu ◽  
Yanli Lv ◽  
Zhenhua Fan ◽  
Weimin Mu ◽  
...  

Introduction: Periductal mastitis (PDM) is a complex benign breast disease with a prolonged course and a high risk of recurrence after treatment. There are many available treatments for PDM, but none is widely accepted. This study aims to evaluate the various treatment failure rates (TFR) of different invasive treatment measures by looking at recurrence and persistence after treatment. In this way, it sets out to inform better clinical decisions in the treatment of PDM. Methods: We searched PubMed, Embase, and Cochrane Library databases for eligible studies about different treatment regimens provided to PDM patients that had been published before October 1, 2019. We included original studies written in English that reported the recurrence and/or persistence rates of each therapy. Outcomes were presented as pooled TFR and 95% CI for the TFR. Results: We included 27 eligible studies involving 1,066 patients in this study. We summarized 4 groups and 10 subgroups of PDM treatments, according to the published studies. Patients treated minimally invasively (group 1) were subdivided into 3 subgroups and pooled TFR were calculated as follows: incision and drainage (n = 73; TFR = 75.6%; 95% CI 27.3–100%), incision alone (n = 74; TFR = 20.1%; 95% CI 0–59.9%), and breast duct irrigation (n = 123; TFR = 19.4%; 95% CI 0–65.0%). Patients treated with a minor excision (excision of the infected tissue and related duct; group 2) were divided into 4 subgroups and pooled TFR were calculated as follows: wound packing alone (n = 127; TFR = 2.1%; 95% CI 0–5.2%), primary closure alone (n = 66; TFR = 37.1%; 95% CI 9.5–64.8%), primary closure under antibiotic treatment cover (n = 55; TFR = 4.8%; 95% CI 0–11.4%) , and additional nipple part removal (n = 232; TFR = 9.6%; 95% CI 5.8–13.4%). Patients treated with a major excision (excision of the infected tissue and the major duct; group 3) included the following 2 subgroups: patients treated with a circumareolar incision (n = 142; TFR = 7.5%; 95% CI 0.4–14.7%) and patients treated with a radial incision of the breast (n = 78; TFR = 0.6%; 95% CI 0–3.6%). Group 4 contained patients receiving different major plastic surgeries. The pooled TFR of this group (n = 86) was 3.4% (95% CI 0–7.5%). Conclusion: Breast duct irrigation, which is the most minimally invasive of all of the treatment options, seemed to yield good outcomes and may be the first-line treatment for PDM patients. Minor excision methods, except for primary closure alone, might be enough for most PDM patients. Major excision, especially with radial incision, was a highly effective salvage therapy. The major plastic surgery technique was also acceptable as an alternative treatment for patients with large lesions and concerns about breast appearance. Incision and drainage and minor excision with primary closure alone should be avoided for PDM patients. Further research is still needed to better understand the etiology and pathogenesis of PDM and explore more effective treatments for this disease.


2021 ◽  
Author(s):  
Prashant More ◽  
Parinita Agarwal ◽  
Pradeep K. Agarwal

Abstract Jatropha, a popular biodiesel crop, suffers severe losses due to Jatropha leaf curl Gujarat virus (JLCuGV) infection in Gujarat (India). Gas chromatography-mass spectrometry (GC-MS) analysis of healthy and infected tissue revealed that the metabolites like sugars, polyols, carboxylic acids, fatty acids, polyphenols, and amino acids were regulated on JLCuGV infection. The sugars (glucose, sucrose, and fructose) increased, while carboxylic acids (malic acid, citric acid and quinic acid) and polyols (galactinol, butanetriol, triethylene glycol, myo-inositol, erythritol) decreased remarkably in infected Jatropha tissue. The increased accumulation of polyphenols in JLCuGV infected tissue suggests activation and involvement of polyphenol biosynthesis pathways as a defence response to geminivirus. All these metabolic variations indicated that sugar metabolism and tricarboxylic acid (TCA) cycle pathways are regulated as a defence response as well as a disease development response to geminivirus infection in Jatropha.


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