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Abstract Immunotherapeutic drugs are promising medicines for cancer treatment. A potential candidate for immunotherapy is interleukin-12 (IL-12), a cytokine well known for its ability to mediate antitumor activity. We developed a plasmid encoding human IL-12 devoid of an antibiotic resistance gene (phIL12). For the approval of phase I clinical trials in basal cell carcinoma (BCC), the regulatory agency requires non-clinical in vivo testing of the pharmacodynamic, pharmacokinetic and toxicological properties of the plasmid. As human IL-12 is not biologically active in mice, a mouse ortholog of the plasmid phIL12 (pmIL12) was evaluated. The evaluation demonstrated the antitumor effectiveness of the protein accompanied by immune cell infiltration. The plasmid was distributed throughout the body, and the amount of plasmid diminished over time in all organs except the skin around the tumor. The therapy did not cause any detectable systemic toxicity. The results of the non-clinical evaluation demonstrated the safety and efficacy of the pmIL12/phIL12 GET, and on the basis of these results, approval was obtained for the initiation of a phase I clinical study in BCC.

Interleukin 12 (IL-12) is a key cytokine that mediates antitumor activity of immune cells. To fulfill its clinical potential, the development is focused on localized delivery systems, such as gene electrotransfer, which can provide localized delivery of IL-12 to the tumor microenvironment. Gene electrotransfer of the plasmid encoding human IL-12 is already in clinical trials in USA, demonstrating positive results in the treatment of melanoma patients. To comply with EU regulatory requirements for clinical application, which recommend the use of antibiotic resistance gene-free plasmids, we constructed and developed the production process for the clinical grade quality antibiotic resistance gene-free plasmid encoding human IL-12 (p21-hIL-12-ORT) and its ortholog encoding murine IL-12 (p21-mIL-12-ORT). To demonstrate the suitability of the p21-hIL-12-ORT or p21-mIL-12-ORT plasmid for the first-in-human clinical trial, the biological activity of the expressed transgene, its level of expression and plasmid copy number were determined in vitro in the human squamous cell carcinoma cell line FaDu and the murine colon carcinoma cell line CT26. The results of the non-clinical evaluation in vitro set the basis for further in vivo testing and evaluation of antitumor activity of therapeutic molecules in murine models as well as provide crucial data for further clinical trials of the constructed antibiotic resistance gene-free plasmid in humans.

In recent years, photosynthetic autotrophic cyanobacteria have attracted interest for biotechnological applications for sustainable production of valuable metabolites. Although biosafety issues can have a great impact on public acceptance of cyanobacterial biotechnology, biosafety of genetically modified cyanobacteria has remained largely unexplored. We set out to incorporate biocontainment systems in the model cyanobacterium Synechocystis sp. PCC 6803. Plasmid-encoded safeguards were constructed using the nonspecific nuclease NucA from Anabaena combined with different metal-ion inducible promoters. In this manner, conditional lethality was dependent on intracellular DNA degradation for regulated autokilling as well as preclusion of horizontal gene transfer. In cells carrying the suicide switch comprising the nucA gene fused to a variant of the copM promoter, efficient inducible autokilling was elicited. Parallel to nuclease-based safeguards, cyanobacterial toxin/antitoxin (TA) modules were examined in biosafety switches. Rewiring of Synechocystis TA pairs ssr1114/slr0664 and slr6101/slr6100 for conditional lethality using metal-ion responsive promoters resulted in reduced growth, rather than cell killing, suggesting cells could cope with elevated toxin levels. Overall, promoter properties and translation efficiency influenced the efficacy of biocontainment systems. Several metal-ion promoters were tested in the context of safeguards, and selected promoters, including a nrsB variant, were characterized by beta-galactosidase reporter assay.

The study was designed to examine the mental health impact of COVID-19 infection in postpartum women and its effects on mother-infant bonding during the first eight weeks postpartum. Fifty-seven consenting eligible postpartum women were recruited for the study. They were assessed at two time points using standardized rating scales to measure distress and uniquely designed scales assessing COVID-19-specific outcome fears and bonding. Almost half [42%] of postpartum women with COVID-19 suffered from a probable anxiety disorder, and one-third [33.3%] suffered from probable depression. The overwhelming majority [91.2%] experienced COVID-19-specific fear. There was an inverse relationship between one dimension of maternal caregiving and self-report depression and anxiety scores, respectively. Additionally, despite discharge, 25% of the mothers had not breastfed the infants till the 8th-week postpartum period, which is in discordance with the World Health Organization (WHO) recommendation of exclusive breastfeeding up to 6 months of age that is widely practiced in India. The novel COVID-19 pandemic was associated with anxiety and depression, impacting mother-infant bonding. Therefore, there is a need for specialized mental health services and individualized breastfeeding interventions for this vulnerable population to ensure positive outcomes., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Aim: To evaluate and correlate the expression of heat shock protein 27 (HSP27) in oral epithelial dysplasia (ED) and oral squamous cell carcinoma (OSCC)., Materials and Methods: This immunohistochemical study of HSP27 expression was performed on 45 samples retrieved from the departmental archives. It included 15 cases of oral ED, 15 cases of OSCC and 15 cases of epithelial hyperplasia (EH). The staining intensity and distribution were scored. The expression was compared between the study groups. Kruskal-Wallis Test, Mann-Whitney U Test and Chi-square tests were performed for statistical analysis using SPSS v21.0 (IBM Corp., Armonk, NY, USA).  Results: There was a statistically significant difference in HSP27 staining parameters between EH, oral ED and OSCC. There was no significant difference between oral ED and OSCC.  Conclusion: HSP27 expression shows enhanced expression in oral ED and OSCC. Its expression should be investigated using larger sample sizes with clinico-pathological correlation to prove its efficiency as a prognostic marker. It will help us in defining treatment modalities so that mortality and morbidity associated with OSCC could be reduced., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Ramalingam et al.)

Background: Myofibroblasts are thought to play critical roles in inflammation, growth, repair, premalignancy, and malignancy. This study was done to evaluate, compare and co- relate the progressive increase in the immunohistochemical expression of myofibroblasts in normal oral mucosa, epithelial dysplasia, and oral squamous cell carcinoma (OSCC). To compare and co-relate the expression of myofibroblasts in normal oral mucosa, epithelial dysplasia, and oral squamous cell carcinoma. To co-relate the progressive increase in myofibroblasts expression in normal oral mucosa, epithelial dysplasia, and oral squamous cell carcinoma., Materials and Method: Forty-nine paraffin-embedded tissue blocks with 7 cases of normal oral mucosa, 21 cases of epithelial dysplasia, and 21 diagnosed cases of OSCCs were studied. The samples were subjected to heat-induced antigen retrieval methods followed by staining using primary mouse monoclonal antibodies against α-smooth muscle actin (SMA) and vimentin. Staining index of all the sections was calculated. Statistical analysis was performed using the Kruskal-Wallis test, Mann-Whitney U test, and Chi-square test. Values of P less than or equal to 0.05 ( P ≤ 0.05) were considered statistically significant., Results: Statistically significant staining index was obtained by α-SMA and vimentin between normal oral mucosa, epithelial dysplasia, and OSCC., Conclusion: Myofibroblast may play a role only during initial tumorigenesis that is the conversion of severe dysplasia into OSCC., Competing Interests: There are no conflicts of interest., (Copyright: © 2020 Journal of Family Medicine and Primary Care.)

Wernicke's encephalopathy (WE) is an uncommon neurological complication in pregnancies complicated with hyperemesis due to thiamine deficiency. In women with hyperemesis, inadvertent glucose administration prior to thiamine supplementation triggers the development of neurological manifestations. Delay in the diagnosis can lead to maternal morbidity, and in one-third of cases may lead to persistence of some neurological deficit. With early recognition and thiamine supplementation, complete recovery is reported. We report a case of WE complicating a case of triplet pregnancy with hyperemesis gravidarum, which highlights the importance of early recognition and treatment, resulting in complete recovery as in the index case., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2019. No commercial re-use. See rights and permissions. Published by BMJ.)

Aim: To describe the ultrasonographic appearance of fetal ventricular aneurysm., Methods: Ultrasound characteristics of two cases of prenatally diagnosed fetal ventricular aneurysm were reviewed., Results: There was one case of left ventricular aneurysm diagnosed by prenatal ultrasound at 38 weeks of gestation. Another case of fetal right ventricular aneurysm was diagnosed prenatally at 36 weeks of gestation. Both the babies expired in the immediate post-natal period., Conclusion: Congenital ventricular aneurysm is a rare cardiac malformation characterized by protrusion or out pouching of a portion of the ventricular wall. It is important to differentiate an aneurysm from diverticulum, as the latter has favorable prognosis. The aneurysms are usually large and have broad-based communication with the ventricular cavity in contrast to diverticuli, which are small and connected by a narrow base.

Chronic kidney disease (CKD) is common after hematopoietic cell transplantation (HCT). We prospectively measured the urinary albumin:creatinine ratio (ACR) in 142 patients. Total (intact) monomeric albumin was determined by liquid chromatography of untreated urine samples collected weekly to day 100 after HCT. Albuminuria was defined as ACR (mg/g creatinine) > 30; proteinuria, as ACR >300. Cox and logistic regression analyses evaluated ACR as a risk factor for clinical events. The prevalence of albuminuria was 37% at baseline, 64% at day 100, and 50% at 1 year. Proteinuria occurred in 4% of patients at baseline, in 15% at day 100, and in 4% at 1 year. Characteristics associated with albuminuria include age, sex, donor type, hypertension, and sinusoidal obstruction syndrome (SOS). Albuminuria was associated with an increased risk of acute graft-versus-host disease (aGVHD) and bacteremia, but not acute kidney injury (AKI). Albuminuria at day 100 was associated with CKD at 1 year (odds ratio = 4.0; 95% confidence interval [CI] = 1.1 to 14.6). Nonrelapse mortality (NRM) risk was elevated (hazard ratio = 6.8; 95% CI = 1.1 to 41.5) in patients with overt proteinuria at day 100. Albuminuria occurs frequently after HCT and is correlated with aGVHD, bacteremia, hypertension, and progression of renal disease. Proteinuria at day 100 is associated with an 6-fold increased risk of NRM by 1 year after HCT.

Background and Objectives: Hematopoietic cell transplantation is a common treatment option for a variety of hematopoietic malignancies. As a result of the use of total body irradiation and/or chemotherapeutic agents, renal dysfunction often ensues. Many pharmacologic agents, such as cyclosporine and high-intensity conditioning regimens, have been linked with thrombotic microangiopathy. In addition, an association between membranous nephropathy and graft-versus-host disease has been reported in this clinical setting., Design, Setting, Participants, and Measurements: A study of autologous and allogeneic hematopoietic cell transplantation patients with renal dysfunction was conducted to document the spectrum of renal manifestations. The pathology files at the University of Washington and University of Chicago Medical Centers were reviewed, and 20 patients with a kidney biopsy after hematopoietic cell transplantation were identified. The histologic findings were correlated with relevant clinical information., Results: A wide spectrum of renal diseases could be classified into four categories: (1) Complications related to hematopoietic cell transplantation (conditioning regimen, immunosuppression, or posttransplantation complications), (2) podocytopathy, (3) membranous nephropathy, or (4) recurrence or persistence of original hematologic disease. Pathologic diagnoses included thrombotic microangiopathy, polyoma virus nephropathy, acute kidney injury/acute tubular necrosis, acute and chronic interstitial nephritis, minimal-change disease, "tip" variant of focal segmental glomerulosclerosis, membranous nephropathy, amyloidosis, and myeloma cast nephropathy. Membranous nephropathy, minimal-change disease, and amyloidosis were common causes of severe proteinuria. Because of the conditioning regimens, posttransplantation complications, and potential nephrotoxic agents used during hematopoietic cell transplantation, it was difficult to attribute the subsequent renal dysfunction to specific factors., Conclusions: The renal biopsy remains essential for diagnosing the underlying injury that can affect one or more compartments of the kidney in this unique clinical setting.