High-Performance Liquid Chromatography (HPLC) is an essential analytical technique in the biopharmaceutical industry, crucial for the separation, identification, and quantification of complex biological molecules such as monoclonal antibodies and recombinant proteins. It plays a vital role in assessing the purity, potency, and stability of biopharmaceutical products, which are critical for regulatory approval. HPLC offers high resolution and sensitivity, allowing for the detection of small quantities of compounds in complex samples. Its versatility is evident in various modes, including reversed-phase, ion-exchange, size-exclusion, and affinity chromatography. However, challenges remain, such as selecting the appropriate stationary phase, addressing peak overlapping and matrix interference, and optimizing operational parameters like flow rate and mobile phase composition. Standardization and method validation are essential for ensuring reproducibility, accuracy, and regulatory compliance in HPLC analyses. The need for reliable reference materials and calibration methods is also a significant challenge. Recent advancements in HPLC technology, including ultra-high-performance liquid chromatography (UHPLC) and hybrid systems that integrate HPLC with mass spectrometry, are helping to overcome these challenges by enhancing sensitivity, resolution, and analysis speed. In summary, as biopharmaceutical products grow more complex, HPLC’s role will continue to evolve, highlighting the need for ongoing research and development to refine this critical analytical tool.

High-Performance Liquid Chromatography (HPLC) is an essential analytical technique in the biopharmaceutical industry, crucial for the separation, identification, and quantification of complex biological molecules such as monoclonal antibodies and recombinant proteins. It plays a vital role in assessing the purity, potency, and stability of biopharmaceutical products, which are critical for regulatory approval. HPLC offers high resolution and sensitivity, allowing for the detection of small quantities of compounds in complex samples. Its versatility is evident in various modes, including reversed-phase, ion-exchange, size-exclusion, and affinity chromatography. However, challenges remain, such as selecting the appropriate stationary phase, addressing peak overlapping and matrix interference, and optimizing operational parameters like flow rate and mobile phase composition. Standardization and method validation are essential for ensuring reproducibility, accuracy, and regulatory compliance in HPLC analyses. The need for reliable reference materials and calibration methods is also a significant challenge. Recent advancements in HPLC technology, including ultra-high-performance liquid chromatography (UHPLC) and hybrid systems that integrate HPLC with mass spectrometry, are helping to overcome these challenges by enhancing sensitivity, resolution, and analysis speed. In summary, as biopharmaceutical products grow more complex, HPLC’s role will continue to evolve, highlighting the need for ongoing research and development to refine this critical analytical tool.

High-Performance Liquid Chromatography (HPLC) is an essential analytical technique in the biopharmaceutical industry, crucial for the separation, identification, and quantification of complex biological molecules such as monoclonal antibodies and recombinant proteins. It plays a vital role in assessing the purity, potency, and stability of biopharmaceutical products, which are critical for regulatory approval. HPLC offers high resolution and sensitivity, allowing for the detection of small quantities of compounds in complex samples. Its versatility is evident in various modes, including reversed-phase, ion-exchange, size-exclusion, and affinity chromatography. However, challenges remain, such as selecting the appropriate stationary phase, addressing peak overlapping and matrix interference, and optimizing operational parameters like flow rate and mobile phase composition. Standardization and method validation are essential for ensuring reproducibility, accuracy, and regulatory compliance in HPLC analyses. The need for reliable reference materials and calibration methods is also a significant challenge. Recent advancements in HPLC technology, including ultra-high-performance liquid chromatography (UHPLC) and hybrid systems that integrate HPLC with mass spectrometry, are helping to overcome these challenges by enhancing sensitivity, resolution, and analysis speed. In summary, as biopharmaceutical products grow more complex, HPLC’s role will continue to evolve, highlighting the need for ongoing research and development to refine this critical analytical tool.

Targeting multiple immune mechanisms may overcome therapy resistance and further improve cancer immunotherapy for humans. Here, we describe the application of virus-like vesicles (VLV) for delivery of three immunomodulators alone and in combination, as a promising approach for cancer immunotherapy. VLV vectors were designed to deliver single chain interleukin (IL)-12, short-hairpin RNA (shRNA) targeting programmed death ligand 1 (PD-L1), and a dominant-negative form of IL-17 receptor A (dn-IL17RA) as a single payload or as a combination payload. Intralesional delivery of the VLV vector expressing IL-12 alone, as well as the trivalent vector (designated CARG-2020) eradicated large established tumors. However, only CARG-2020 prevented tumor recurrence and provided long-term survival benefit to the tumor-bearing mice, indicating a benefit of the combined immunomodulation. The abscopal effects of CARG-2020 on the non-injected contralateral tumors, as well as protection from the tumor cell re-challenge, suggest immune-mediated mechanism of protection and establishment of immunological memory. Mechanistically, CARG-2020 potently activates Th1 immune mechanisms and inhibits expression of genes related to T cell exhaustion and cancer-promoting inflammation. The ability of CARG-2020 to prevent tumor recurrence and to provide survival benefit makes it a promising candidate for its development for human cancer immunotherapy.

Background: Face masks have become an important tool for preventing the spread of respiratory diseases. However, we hypothesized that face masks with reduced nasal airflow may alter pulmonary artery systolic pressure (PASP).Purpose: This study aimed to evaluate the effect of face masks on PASP in children and adolescents. Methods: This case-control study was conducted between March 2021 and April 2022 at the Pediatric Cardiovascular Research Center in Isfahan, Iran. Using a convenience sampling method, a total of 120 children and adolescents, boys and girls aged 3–18 years, were allocated into 2 groups of 60 each (case group with congenital heart disease (CHD), control group of healthy subjects). For each patient in the case and control groups, echocardiography (ECHO), heart rate (HR), and blood oxygen saturation (SpO2) were performed and measured twice—once with a surgical mask and once without a surgical mask—by a pediatric cardiologist at 10-min intervals. Results: A total of 110 participants were analyzed. The mean patient age was 9.58±3.40 years versus 10.20±4.15 years in the case (n=50) and control (n=60) groups, respectively. Approximately 76.0% (n=38) of the participants in the case group versus 60.0% of those in the control group were male. In the case and control groups, there was a statistically significant reduction in the mean changes in tricuspid regurgitation (P=0.001), pulmonary regurgitation (P=0.002), and PASP (P=0.001) after face mask removal. Although this study showed a reduction in pulmonary arterial pressure after face mask removal in patients with CHD and healthy subjects, no significant changes in HR (P=0.535) or SpO2 (P=0.741) were observed in either group. Conclusion Wearing a face mask increased PASP in healthy children and adolescents with CHD; however, the SPO2 and HR remained unchanged. Therefore, mask removal during ECHO is recommended.

Background: Face masks have become an important tool for preventing the spread of respiratory diseases. However, we hypothesized that face masks with reduced nasal airflow may alter pulmonary artery systolic pressure (PASP).Purpose: This study aimed to evaluate the effect of face masks on PASP in children and adolescents. Methods: This case-control study was conducted between March 2021 and April 2022 at the Pediatric Cardiovascular Research Center in Isfahan, Iran. Using a convenience sampling method, a total of 120 children and adolescents, boys and girls aged 3–18 years, were allocated into 2 groups of 60 each (case group with congenital heart disease (CHD), control group of healthy subjects). For each patient in the case and control groups, echocardiography (ECHO), heart rate (HR), and blood oxygen saturation (SpO2) were performed and measured twice—once with a surgical mask and once without a surgical mask—by a pediatric cardiologist at 10-min intervals. Results: A total of 110 participants were analyzed. The mean patient age was 9.58±3.40 years versus 10.20±4.15 years in the case (n=50) and control (n=60) groups, respectively. Approximately 76.0% (n=38) of the participants in the case group versus 60.0% of those in the control group were male. In the case and control groups, there was a statistically significant reduction in the mean changes in tricuspid regurgitation (P=0.001), pulmonary regurgitation (P=0.002), and PASP (P=0.001) after face mask removal. Although this study showed a reduction in pulmonary arterial pressure after face mask removal in patients with CHD and healthy subjects, no significant changes in HR (P=0.535) or SpO2 (P=0.741) were observed in either group. Conclusion Wearing a face mask increased PASP in healthy children and adolescents with CHD; however, the SPO2 and HR remained unchanged. Therefore, mask removal during ECHO is recommended.

Background: Face masks have become an important tool for preventing the spread of respiratory diseases. However, we hypothesized that face masks with reduced nasal airflow may alter pulmonary artery systolic pressure (PASP).Purpose: This study aimed to evaluate the effect of face masks on PASP in children and adolescents. Methods: This case-control study was conducted between March 2021 and April 2022 at the Pediatric Cardiovascular Research Center in Isfahan, Iran. Using a convenience sampling method, a total of 120 children and adolescents, boys and girls aged 3–18 years, were allocated into 2 groups of 60 each (case group with congenital heart disease (CHD), control group of healthy subjects). For each patient in the case and control groups, echocardiography (ECHO), heart rate (HR), and blood oxygen saturation (SpO2) were performed and measured twice—once with a surgical mask and once without a surgical mask—by a pediatric cardiologist at 10-min intervals. Results: A total of 110 participants were analyzed. The mean patient age was 9.58±3.40 years versus 10.20±4.15 years in the case (n=50) and control (n=60) groups, respectively. Approximately 76.0% (n=38) of the participants in the case group versus 60.0% of those in the control group were male. In the case and control groups, there was a statistically significant reduction in the mean changes in tricuspid regurgitation (P=0.001), pulmonary regurgitation (P=0.002), and PASP (P=0.001) after face mask removal. Although this study showed a reduction in pulmonary arterial pressure after face mask removal in patients with CHD and healthy subjects, no significant changes in HR (P=0.535) or SpO2 (P=0.741) were observed in either group. Conclusion Wearing a face mask increased PASP in healthy children and adolescents with CHD; however, the SPO2 and HR remained unchanged. Therefore, mask removal during ECHO is recommended.

Background: Face masks have become an important tool for preventing the spread of respiratory diseases. However, we hypothesized that face masks with reduced nasal airflow may alter pulmonary artery systolic pressure (PASP).Purpose: This study aimed to evaluate the effect of face masks on PASP in children and adolescents. Methods: This case-control study was conducted between March 2021 and April 2022 at the Pediatric Cardiovascular Research Center in Isfahan, Iran. Using a convenience sampling method, a total of 120 children and adolescents, boys and girls aged 3–18 years, were allocated into 2 groups of 60 each (case group with congenital heart disease (CHD), control group of healthy subjects). For each patient in the case and control groups, echocardiography (ECHO), heart rate (HR), and blood oxygen saturation (SpO2) were performed and measured twice—once with a surgical mask and once without a surgical mask—by a pediatric cardiologist at 10-min intervals. Results: A total of 110 participants were analyzed. The mean patient age was 9.58±3.40 years versus 10.20±4.15 years in the case (n=50) and control (n=60) groups, respectively. Approximately 76.0% (n=38) of the participants in the case group versus 60.0% of those in the control group were male. In the case and control groups, there was a statistically significant reduction in the mean changes in tricuspid regurgitation (P=0.001), pulmonary regurgitation (P=0.002), and PASP (P=0.001) after face mask removal. Although this study showed a reduction in pulmonary arterial pressure after face mask removal in patients with CHD and healthy subjects, no significant changes in HR (P=0.535) or SpO2 (P=0.741) were observed in either group. Conclusion Wearing a face mask increased PASP in healthy children and adolescents with CHD; however, the SPO2 and HR remained unchanged. Therefore, mask removal during ECHO is recommended.

Aims: This study was aimed to monitor the asymptomatic carriage of coagulase-positive staphylococcal bacteria among university male students and detect the prevalence of virulence marker genes that encode methicillin resistance (mecA) and Panton-Valentine leukocidin (PVL) toxin among the isolates. Methodology and results: Single nasal swaps were collected from 144 participating students who resided at four different locations within Al-Madinah city. A total of 112 Gram-positive staphylococcal isolates were recovered from the 144 participants (carriage rate of 77.8%). Coagulase-positive staphylococci were differentiated using duplex PCR amplification of the 16S rRNA and nuc genes and accounted for 30 isolates (carriage rate of 20.8%). These isolates were most prevalent in the northern and southern parts of Al-Madinah city, while the lowest numbers of isolates were detected in students of the eastern part. Coagulase-positive isolates were further phenotypically characterized for methicillin resistance by the disc diffusion method. Uniplex PCR assays were conducted to screen for mecA- and PVL toxin-encoding genes. The mecA gene was amplified from all 15 (50%) methicillin-resistant coagulase-positive isolates, while the PVL toxin-encoding gene was detected in 19 isolates (63.3%), 10 (33.3%) of which contained the mecA gene. Lastly, PCR amplification of the NRPS gene from coagulase-positive isolates revealed the absence of Staphylococcus argenteus, the recently discovered genetically divergent lineage of Staphylococcus aureus. Conclusion, significance and impact of study An elevated prevalence of coagulase-positive isolates harboring mecA and PVL virulence genes was observed compared with previous investigations. This poses a potential threat if they spread among the population, resulting in outbreaks of community-acquired infections.

Shiga toxin-producing Escherichia coli is the most prevalent bacterial strain responsible for Shiga toxin-related infections. While Shiga toxin is inherently toxic, it has potential therapeutic applications as a component of anticancer drugs. Despite its association with infections and harmful effects on human health, Shiga toxin is being explored as a viable element in drug delivery systems targeting cancer cells. The findings indicate that the production of mutated bacteria containing Shiga toxin is an effective preventive strategy for immunization against these toxins. Furthermore, the B subunit of Shiga toxin shows promise for imaging cancer cells, opening new paths for therapeutic interventions.