Identification of pathological subtypes of early lung adenocarcinoma based on artificial intelligence parameters and CT signs

Objective: To explore the value of quantitative parameters of artificial intelligence and computed tomography (CT) signs in identifying pathological subtypes of lung adenocarcinoma appearing as ground-glass nodules (GGNs). Methods: CT images of 224 GGNs from 210 individuals were collected retrospectively and pathologically classified into atypical adenomatous hyperplasia (AAH)/adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA), and invasive adenocarcinoma (IAC) groups. Artificial intelligence was used to identify GGNs and to obtain quantitative parameters, and CT signs were recognized manually. The mixed predictive model based on logistic multivariate regression was evaluated. Results: Of the 224 GGNs, 55, 93, and 76 were AAH/AIS, MIA, IAC, respectively. In terms of artificial intelligence parameters, from AAH/AIS to MIA, and IAC, there was a gradual increase in two-dimensional mean diameter, three-dimensional mean diameter, mean CT value, maximum CT value, and volume of GGNs (all P < 0.0001). Except for the CT signs of the location, and the tumor-lung interface, there were significant differences among the three groups in the density type, shape, vacuole signs, air bronchogram, lobulation, spiculation, pleural indentation, and vascular convergence signs (all P < 0.05). The areas under the curve (AUC) of predictive model 1 for identifying the AAH/AIS and MIA and model 2 for identifying MIA and IAC were 0.779 and 0.918, respectively, which were greater than the quantitative parameters independently (all P < 0.05). Conclusion: Artificial intelligence parameters are valuable for identifying subtypes of early lung adenocarcinoma, and when combined with CT signs to improve its diagnostic efficacy.

CT-Assisted Improvements in the Accuracy of the Intraoperative Frozen Section Examination of Ground-Glass Density Nodules

Objectives. The intraoperative frozen section examination (IFSE) of pulmonary ground-glass density nodules (GGNs) is a great challenge. In the present study, through comparing the correlation between the computed tomography (CT) findings and pathological diagnosis of GGNs, the CT features as independent risk factors affecting the examination were defined, and their value in the rapid intraoperative examination of GGNs was explored. Methods. The relevant clinical data of 90 patients with GGNs on CT were collected, and all CT findings of GGNs, including the maximum transverse diameter, average CT value, spiculation, solid component, vascular sign, air sign, bronchus sign, lobulation, and pleural indentation, were recorded. All the cases received thoracoscopic surgery, and final pathological results were obtained. The cases were divided into three groups on the basis of pathological diagnosis: benign/atypical adenomatous hyperplasia (AAH), adenocarcinoma in situ (AIS)/microinvasive adenocarcinoma (MIA), and invasive adenocarcinoma (IAC). The CT findings were analyzed statistically, the independent risk factors were identified through the intergroup bivariate logistic regression analysis on variables with statistically significant differences, and a receiver operating curve (ROC) was plotted to establish a logistic regression model for diagnosing GGNs. A retrospective analysis was conducted on the coincidence rate of the rapid intraoperative and routine postoperative pathological examinations of the 90 cases with GGNs. The relevant clinical data of 49 cases with GGNs were collected. Conventional rapid intraoperative examination and CT-assisted rapid intraoperative examination were performed, and their coincidence rates with routine postoperative pathological examinations were compared. Results. No statistical differences in the onset age, gender, smoking history, and family history of malignant tumors were found among cases with GGNs in the identification of benign/AAH, AIS/MIA, and IAC ( P = 0.158 , P = 0.947 , P = 0.746 , P = 0.566 ). No statistically significant difference was found among the three groups in terms of CT findings, such as lobulation, bronchus sign, pleural indentation, spiculation, vascular sign, and solid component ( P > 0.05 ). The air sign, the maximum transverse diameter of GGNs, and average CT value showed statistically significant differences among the groups ( P < 0.001 , P < 0.05 , P < 0.001 ). Bivariate logistic regression analysis was performed on three risk factors, and the predicted probability value was obtained. A ROC curve was plotted by using the maximum transverse diameter as a predictor for analysis between the groups with benign/AAH and AIS/MIA, and the results demonstrated that the area under the curve (AUC) was 0.692. A ROC curve was plotted by using the predicted probability value, maximum transverse diameter, and average CT value as predictors for distinguishing between the groups with AIS/MIA and IAC, and the results showed that the AUC values of the predicted probability value, maximum transverse diameter, and CT value were 0.920, 0.816, and 0.772, respectively. A regression model Logit P = 2.304 − 2.689 X 1 + 0.302 X 2 + 0.011 X 3 was established to identify GGNs as IAC, obtaining AUC values of up to 0.920 for the groups with AIS/MIA and IAC, the sensitivity of 0.821, and the specificity of 0.894. The coincidence rate of rapid intraoperative and routine postoperative pathological examinations taken for modeling was 79.3%, that of conventional IFSE and postoperative pathological examination in prospective studies was 83.7%, and that of CT-assisted rapid intraoperative and postoperative pathological examinations was 98.0%. The former two were statistically different from the last one ( P = 0.003 and P = 0.031 , respectively). Conclusion. The air sign, maximum transverse diameter, and average CT value of the CT findings of GGNs had superior capabilities to enhance the pathologic classification of GGNs. The auxiliary function of the comprehensive multifactor analysis of GGNs was better than that of single-factor analysis. CT-assisted diagnosis can improve the accuracy of rapid intraoperative examination, thereby increasing the accuracy of the selection of operative approaches in clinical practice.

Comparative Diagnostic Accuracy of Mean and Lowest Cycle Threshold Levels in Xpert MTB/RIF Assay and Molecular Epidemiology of Missing Probes In Rifampicin Resistant Tubercular Cases From A Tertiary Care Hospital

Background: The comparative diagnostic accuracy of mean and lowest Ct values needs to be evaluated for the assessment of mycobacterial burden in tubercular cases. Mutation in any codon of 81 base-pair core regions prevents the hybridization of one or more of five overlapping Probes A-Ein Xpert MTB/RIF assay indicated by “missing probe. Molecular epidemiology of missing probes may prove useful in tracing the source of infection and selection of a more suitable drug regimen for treatment. Methods: This study included 65 rifampicin resistant cases and an equal number of rifampicin sensitive cases detected by Xpert MTB/RIF assay. Only samples tested positive for tubercular bacilli were included. The information regarding the tubercular load, Ct values of five probes targeting the rpoB gene, lowest Ct value among the five probes, missing probe in rifampicin resistant cases and time taken for the entire cycle were recorded in each case. Results: Lowest Ct is a stronger indicator of tubercular load than the mean Ct value. E probe was found to be missing in majority (64.6%) of the cases, followed by A (6.2%), B and D (4.6%), C (1.5%) probes. In 7.6% cases, more than one probe was missing. None of the probe was missing in 10.6% of rifampicin resistant cases. Conclusions: Lowest Ct value was found to be a better tool than mean Ct value for the determination of mycobacterium burden. Molecular epidemiology of missing probes could be useful in the development of new probes for the detection of rifampicin resistance.

Performance of Rapid Antigen Tests for COVID-19 Diagnosis: A Systematic Review and Meta-Analysis

The identification of viral RNA using reverse transcription quantitative polymerase chain reaction (RT-qPCR) is the gold standard for identifying an infection caused by SARS-CoV-2. The limitations of RT-qPCR such as requirement of expensive instruments, trained staff and laboratory facilities led to development of rapid antigen tests (RATs). The performance of RATs has been widely evaluated and found to be varied in different settings. The present systematic review aims to evaluate the pooled sensitivity and specificity of the commercially available RATs. This review was registered on PROSPERO (registration number: CRD42021278105). Literature search was performed through PubMed, Embase and Cochrane COVID-19 Study Register to search studies published up to 26 August 2021. The overall pooled sensitivity and specificity of RATs and subgroup analyses were calculated. Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) was used to assess the risk of bias in each study. The overall pooled sensitivity and specificity of RATs were 70% (95% CI: 69–71) and 98% (95% CI: 98–98), respectively. In subgroup analyses, nasal swabs showed the highest sensitivity of 83% (95% CI: 80–86) followed by nasopharyngeal swabs 71% (95% CI: 70–72), throat swabs 69% (95% CI: 63–75) and saliva 68% (95% CI: 59–77). Samples from symptomatic patients showed a higher sensitivity of 82% (95% CI: 82–82) as compared to asymptomatic patients at 68% (95% CI: 65–71), while a cycle threshold (Ct) value ≤25 showed a higher sensitivity of 96% (95% CI: 95–97) as compared to higher Ct value. Although the sensitivity of RATs needs to be enhanced, it may still be a viable option in places where laboratory facilities are lacking for diagnostic purposes in the early phase of disease.

Challenges in Laboratory Diagnosis of Malaria in a Low-Resource Country at Tertiary Care in Eastern Nepal: A Comparative Study of Conventional vs. Molecular Methodologies

For ongoing malaria elimination programmes, available methods such as microscopy and rapid diagnostic tests (RDTs) cannot detect all malaria cases in acute febrile illness. These methods are entirely dependent on the course of infection, parasite load, and skilled technical resources. Our study objectives were to estimate the performance of light microscopy and a RDT as well as real-time PCR for the detection of the Plasmodium parasite. Altogether, 52 blood samples collected from patients with acute febrile illness were tested by microscopy, RDT, and real-time PCR. The results were compared in terms of sensitivity and specificity. Microscopy detected the malaria parasite in 5.8% of the blood samples whereas 13.5% were detected by the RDT and 27% by real-time PCR. Considering real-time PCR as the gold standard method, microscopy had a sensitivity of 21.4% and a specificity of 100%, and the RDT had a sensitivity of 28.6% and a specificity of 92.1%. Microscopy together with the RDT successfully detected malaria positive cases in blood samples of Ct value below 20, but both were unable to detect malaria cases between 26–40 Ct value ranges amplified by real-time PCR. Despite various diagnostic tools being available, microscopy still remains the method of choice for diagnosis, while the RDT is user-friendly when applied at the point of care. However, our preliminary results emphasize the need to implement the test with higher sensitivity and specificity in the context of a malaria elimination programme. Such programmes can be a crucial opportunity to understand the species prevalent in a low-endemic region. However, these results should be further verified with a large cohort study to document the submicroscopic infection.

Characteristics of Lumbar Bone Density in Middle-Aged and Elderly Subjects: A Correlation Study between T-Scores Determined by the DEXA Scan and Hounsfield Units from CT

Purpose. To describe the characteristics of lumbar bone density in middle-aged and elderly subjects and explore whether there is a correlation between computed tomography (CT) values and the bone mineral density (BMD) T-scores of the lumbar vertebral cancellous bone. Methods. Forty-two subjects, including 25 males and 17 females, with a mean age of 56 years, who underwent BMD measurement and lumbar multislice spiral CT scan at the China Rehabilitation Research Center from January 2019 to December 2019 were selected. Dual-energy X-ray absorptiometry (DEXA) was applied to obtain the total BMD T-scores of the lumbar L1–L4 vertebrae. Results. The CT values decreased from L1 to L4 and were 145.91 ± 8.686 HU, 143.18 ± 8.598 HU, 137.39 ± 8.276 HU, and 135.23 ± 8.219 HU, respectively. The total CT value of L1–L4 was 140.43 ± 4.199 HU. The mean total BMD T-score of L1–L4 was −0.94. The CT values of the L1–L4 vertebrae were positively correlated with the total BMD T-scores of L1–L4 (r = 0.349, P < 0.001 ). The CT value of the left third of the same vertebrae was the highest, and there was a strong positive correlation between the regional CT value of the lumbar spine and the entire vertebra CT values (r > 0.7). Conclusion. The CT values of the lumbar spine can assist the measurement of the T-scores of lumbar BMD, which could aid in early opportunistic screening for osteopenia and preventing osteoporosis and vertebral compression fractures in middle-aged and elderly subjects. This trial is registered with ChiCTR2100049571.

SARS-CoV-2 Viral Load Assessment in Lung Transplantation

In the era of COVID-19 pandemic, organ transplantation programs were facing serious challenges. The lung transplantation donor pool was extremely limited and SARS-CoV-2 viral load assessment has become a crucial part of selecting an optimal organ donor. Since COVID-19 is a respiratory disease, the viral load is thought to be more important in lung transplantations as compared to other solid organ transplantations. We present two challenging cases of potential lung donors with a questionable COVID-19 status. Based on these cases, we suggest that the cycle threshold (Ct) value should always be requested from the laboratory and the decision whether to proceed with transplantation should be made upon complex evaluation of diverse criteria, including the nasopharyngeal swab and bronchoalveolar lavage PCR results, the Ct value, imaging findings and the medical history. However, as the presence of viral RNA does not ensure infectivity, it is still to be clarified which Ct values are associated with the viral viability. Anti-SARS-CoV-2 IgA antibodies may support the diagnosis and moreover, novel methods, such as quantifying SARS-CoV-2 nucleocapsid antigen in serum may provide important answers in organ transplantations and donor selections.

Oral antiviral clevudine compared with placebo in Korean COVID-19 patients with moderate severity

Background: Clevudine, an antiviral drug for chronic hepatitis B virus infection, is expected to inhibit the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. Therefore, we conducted a prospective, single-blind, proof of concept clinical study to examine the antiviral efficacy and safety of clevudine compared to placebo in Korean corona virus disease 19 (COVID-19) patients with moderate severity. Methods: Adults with confirmed SARS-CoV-2 infection and symptom onset within 7 days were randomized 2:1 to 120 mg clevudine or placebo to receive one of treatments orally once-daily for 14 days. Antiviral efficacy outcomes were the proportion of patients with real-time reverse transcription polymerase chain reaction (RT-PCR) negative result for SARS-CoV-2 infection and cycle threshold (Ct) value changes from baseline. Clinical efficacy outcomes included proportion of patients who showed improvement in lung involvement by imaging tests, proportion of patients with normal body temperature, proportion of patients with normal oxygen saturation, and the changes in C-reactive protein (CRP) from baseline. Safety outcomes included changes in clinical laboratory tests, vital signs measurement, and physical examination from baseline, and incidence of adverse events. Results: The proportion of patients with real-time RT-PCR negative test and Ct value changes showed no significant difference between clevudine group and placebo group. The changes in Ct value from baseline were significantly greater in clevudine group compared to placebo group in patients with hypertension, and patients who underwent randomization during the first 5 and 7 days after the onset of symptoms. All clinical efficacy outcomes had no significant difference between clevudine group and placebo group. Clevudine was well tolerated and there was no significant difference in safety profile between two treatment groups. Conclusions: This is the first clinical study to compare the antiviral efficacy and safety of clevudine to placebo in Korean COVID-19 patients with moderate severity. The study has demonstrated a possible favorable outcome for the reduction of SARS-CoV-2 replication, with acceptable safety profile, when COVID-19 patients were treated with clevudine. Further large-scale clinical studies, preferably with various clinical endpoints and virus titer evaluation, are required to better understand the effectiveness of using clevudine in COVID-19 treatment. Considering recent trend in clinical development for antiviral drugs, we need to design a clinical study aiming for reducing clinical risk of COVID-19 in mild to moderate patients with at least one risk factor for serious illness.

Determination of Time of Death Based on Blood Ring Formation and RNA Degradation

Time of death (TOD) determination is crucial in criminal cases. The method used to determine the TOD so far is only based on the state of the corpse found, therefore a new method is needed to improve the accuracy of the TOD determination. This study aims to determine the TOD based on a new method, namely the formation of blood rings and RNA degradation. Blood is commonly found in crime scenes. Blood consists of liquid part that is plasma and cellular part consisting of erythrocytes, leukocyte, and thrombocytes. The composition of blood as a liquid that contains dissolved solids makes the drops of dried blood forming “coffee ring effect”, which is a ring-like form on the perimeter of a blood drop. Coffee ring effect is used as an indicator of time by looking at the thickness of the ring formed from the perimeter of the blood drop to the middle which increases with time. RNA degradation was observed using Peptidylprolyl isomerase A (PPIA) gene. The PPIA gene is found in leukocyte and is used to see the degradation of RNA per 30 minutes period using the RT-PCR and qPCR methods. Degradation was observed by comparing the cycle threshold (ct) value of the standard curve with the ct value of the samples per unit time. TOD could be determined by the percentage of the blood ring thickness up until 120th minutes, and by observing the degradation of RNA until the 60th minute, after that the RNA had completely degraded.

Is the Infection of the SARS-CoV-2 Delta Variant Associated With the Outcomes of COVID-19 Patients?

Background: Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) Delta variant (B.1.617.2) has been responsible for the current increase in Coronavirus disease 2019 (COVID-19) infectivity rate worldwide. We compared the impact of the Delta variant and non-Delta variant on the COVID-19 outcomes in patients from Yogyakarta and Central Java provinces, Indonesia.Methods: In this cross-sectional study, we ascertained 161 patients, 69 with the Delta variant and 92 with the non-Delta variant. The Illumina MiSeq next-generation sequencer was used to perform the whole-genome sequences of SARS-CoV-2.Results: The mean age of patients with the Delta variant and the non-Delta variant was 27.3 ± 20.0 and 43.0 ± 20.9 (p = 3 × 10−6). The patients with Delta variant consisted of 23 males and 46 females, while the patients with the non-Delta variant involved 56 males and 36 females (p = 0.001). The Ct value of the Delta variant (18.4 ± 2.9) was significantly lower than that of the non-Delta variant (19.5 ± 3.8) (p = 0.043). There was no significant difference in the hospitalization and mortality of patients with Delta and non-Delta variants (p = 0.80 and 0.29, respectively). None of the prognostic factors were associated with the hospitalization, except diabetes with an OR of 3.6 (95% CI = 1.02–12.5; p = 0.036). Moreover, the patients with the following factors have been associated with higher mortality rate than the patients without the factors: age ≥65 years, obesity, diabetes, hypertension, and cardiovascular disease with the OR of 11 (95% CI = 3.4–36; p = 8 × 10−5), 27 (95% CI = 6.1–118; p = 1 × 10−5), 15.6 (95% CI = 5.3–46; p = 6 × 10−7), 12 (95% CI = 4–35.3; p = 1.2 × 10−5), and 6.8 (95% CI = 2.1–22.1; p = 0.003), respectively. Multivariate analysis showed that age ≥65 years, obesity, diabetes, and hypertension were the strong prognostic factors for the mortality of COVID-19 patients with the OR of 3.6 (95% CI = 0.58–21.9; p = 0.028), 16.6 (95% CI = 2.5–107.1; p = 0.003), 5.5 (95% CI = 1.3–23.7; p = 0.021), and 5.8 (95% CI = 1.02–32.8; p = 0.047), respectively.Conclusions: We show that the patients infected by the SARS-CoV-2 Delta variant have a lower Ct value than the patients infected by the non-Delta variant, implying that the Delta variant has a higher viral load, which might cause a more transmissible virus among humans. However, the Delta variant does not affect the COVID-19 outcomes in our patients. Our study also confirms that older age and comorbidity increase the mortality rate of patients with COVID-19.