Michael J. Paidas

The number of patients with long COVID continues to rise as COVID-19 persists, yet research on its implications for female reproductive health remains scarce. Long COVID is linked to an increased risk of abnormal uterine bleeding, heavier menstrual flow, prolonged periods, and breakthrough bleeding. We therefore examined whether uterine changes occur in an animal model of long COVID. Female A/J mice were inoculated with murine hepatitis virus strain 1 (MHV-1), a murine coronavirus model used to investigate long COVID-like sequelae, and a subset of infected mice received the synthetic peptide SPIKENET (SPK). Mice infected with murine hepatitis virus strain 1 (MHV-1) showed chronic uterine inflammation and myometrial changes exhibiting fibroid-like features one year post-infection. We also observed changes in α-SMA, desmin, h-caldesmon, and vimentin staining, suggesting a myometrial origin for the lesions. Further, we found pathological changes that include hyperplastic foci in the outer layer of the myometrium, a uterus with intraluminal proteinaceous fluid, and a circumscribed, round-to-oval focus of benign hyperplastic myometrium with spindle nuclei and eosinophilic cytoplasm, with interspersed small, round lymphocytes and plasma cells, consistent with mild chronic inflammation. MHV-1-inoculated mice also showed an oviduct with several lymphocytes located toward the lumen, as well as a uterus with an ectatic lumen, inspissated intraluminal fibrinoid material, and mild-to-moderate subendometrial chronic inflammation (with scattered small lymphocytes). Additionally, immunofluorescence showed increased expression of conventional and unconventional uterine fibroid markers, including PLP1, versican, PR-A, PR-B, MSH2, TGF-β1 and 3, IGF-1, CA125, and HMGA2. Serum biomarker analysis showed increased IGF-1 levels but not increased levels of the other proteins identified by immunofluorescence. Furthermore, histopathology showed no uterine fibroid-like lesions in MHV-1-inoculated mice treated with the synthetic peptide SPK (known to prevent MHV-1 and SARS-CoV-2 infection in animals). Moreover, SPK prevented increases in the above-mentioned fibroid markers as measured by immunofluorescence and partially reduced IGF-1 expression in the serum. These findings suggest that uterine fibroids may develop during long COVID in humans.

Introduction: Trauma is the leading cause of non-obstetrical maternal death and is linked to adverse pregnancy outcomes, including placental abruption, preterm birth, and stillbirth. Although placental injury is a known consequence of maternal trauma, understanding its direct effect on trophoblast function remains limited. This study aims to examine the cellular effects of mechanical trauma on human trophoblasts in vitro . Methods: Human trophoblast cells (HTR-8/SVneo) were cultured and exposed to controlled mechanical trauma using a fluid percussion injury (FPI) model, previously validated in traumatic brain injury research. Cell viability was measured 48 h after injury with a fluorometric assay kit. Immunofluorescence and Western blot analyses were carried out to assess the levels of mitochondrial markers (TOM20, COX1, VDAC1) and the antiangiogenic factor sFlt-1. Results: Trophoblasts subjected to trauma exhibited reduced viability compared with controls. Expression of sFlt-1 and SDHA increased in injured cells, indicating a cellular stress response. Western blot results revealed a twofold decrease in TOM20 levels and a slight reduction in VDAC1 levels, while COX1 levels remained unchanged. Moreover, next-generation sequencing of trophoblasts after trauma showed altered gene expression involving several injury pathways, including mitochondrial stress response, immune dysregulation, cytoskeletal changes, endothelial and angiogenic signaling pathways, calcium signaling, metabolic imbalance, and inflammation. These results suggest that mechanical trauma causes mitochondrial dysfunction and disrupts angiogenic signaling in trophoblasts. Discussion: Trauma induces significant mitochondrial changes and cellular stress, characterized by reduced viability, decreased TOM20 and VDAC1 levels, and increased sFlt-1 and SDHA levels. These patterns resemble those seen in hypoxia-related placental disorders such as preeclampsia, fetal distress, and placental abruption. Understanding these mechanisms may help clarify how maternal trauma contributes to adverse pregnancy outcomes. Further studies using in vivo models are required to explore the clinical relevance of these findings.

Ionizing radiation (IR) damage, whether intentional, accidental, or therapeutic, reverberates throughout the body. The FDA has approved the use of palliative care and leukocyte growth factors for hematologic depletion (H-ARS), but no treatments have been approved for gastrointestinal damage (GI-ARS). Synthetic PIF (sPIF) safely replicates endogenous PIF’s preventative, reparatory, and regenerative effects. Specifically relevant for radiation-induced damage, sPIF reduces oxidative stress. Subcutaneous (SC) sPIF prevents LD100/30 mortality at 2 w post-treatment and restores GI function post-6Gy exposure (in mice). Daily SC sPIF for 14 d, starting 24 h post-LD85/40 (7.17 Gy) total-body ɣ-irradiation (TBI), increases survival 2.76-fold (14.7% (5/34) to 40.7% (24/59); p = 0.0036) by 4 w post-therapy. All sPIF-treated mice survived for the first 10 d, while only 80% survived in the sham group (p = 0.0034). The sPIF-treated group reached 50% survival at 23 di, while this figure was reached at 19 d in the sham-treated group, with a 21% delay. Importantly, sPIF delays weight loss for up to 25 d, increasing weight by ~12% above baseline and improving colon architecture; in the sham, both weight and colon recovery failed. sPIF partially prevents declines in hemoglobin and platelets, with no bleeding/sepsis observed, while WBC counts declined. sPIF’s safety and lack of deleterious drug-to-drug interaction were documented in a First-in-Human, FDA-approved clinical trial for autoimmune disease. Chronic FDA-guided toxicology/toxicokinetic studies demonstrated NOAEL and the highest safety margin. Collectively, sPIF safely and significantly increases survival, weight gain, and colon crypt scale, directly supporting its suitability for FDA-ARS animal-rule approval.

Glucagon-like peptide 1 (GLP-1), a hormone derived from the proglucagon gene, regulates various physiological processes; however, its impact on pregnancy outcomes remains poorly understood. Assessing the effects of GLP-1 on neonates is vital as GLP-1 is increasingly administered during pregnancy. This study evaluates the effect of GLP-1 exposure on maternal complications and neonatal defects in mice. Pregnant female A/J mice received subcutaneous injections of recombinant GLP-1 (rGLP-1; 1000 nmol/kg) on embryonic day 1 (EP, early pregnancy) or day 15 (E15, late pregnancy). Maternal and neonatal body weights, morphology, and mortality were recorded, and mRNA sequencing was conducted to analyze gene expression in neonatal tissues. Maternal body weight decreased following rGLP-1 exposure, and pups born to both the early and late exposure groups experienced significant weight loss. Pups in the late exposure group exhibited uniform skin detachment and a dramatically higher mortality rate than those born to the early exposure group. Further, RT-PCR analysis confirms the significantly increased expression of selected genes in the skin and associated pathogenesis. RNA sequencing of pups’ skin, brain, lung, and liver tissues from the late exposure group showed altered gene expression. Since maternal weight loss, increased neonatal mortality, and altered gene expression have been observed, GLP-1 receptor agonists (GLP-1RAs) should be avoided during pregnancy.

Background: Accumulating evidence indicates that SARS-CoV-2 infection results in long-term multiorgan complications, with the kidney being a primary target. This study aimed to characterize the long-term transcriptomic changes in the kidney following coronavirus infection using a murine model of MHV-1-induced SARS-like illness and to evaluate the therapeutic efficacy of SPIKENET (SPK). Methods: A/J mice were infected with MHV-1. Renal tissues were collected and subjected to immunofluorescence analysis and Next Generation RNA Sequencing to identify differentially expressed genes associated with acute and chronic infection. Bioinformatic analyses, including PCA, volcano plots, and GO/KEGG pathway enrichment, were performed. A separate cohort received SPK treatment, and comparative transcriptomic profiling was conducted. Gene expression profile was further confirmed using real-time PCR. Results: Acute infection showed the upregulation of genes involved in inflammation and fibrosis. Long-term MHV-1 infection led to the sustained upregulation of genes involved in muscle regeneration, cytoskeletal remodeling, and fibrotic responses. Notably, both expression and variability of SLC22 and SLC22A8, key proximal tubule transporters, were reduced, suggesting a loss of segment-specific identity. Further, SLC12A1, a critical regulator of sodium reabsorption and blood pressure, was downregulated and is associated with the onset of polyuria and hydronephrosis. SLC transporters exhibited expression patterns consistent with tubular dysfunction and inflammation. These findings suggest aberrant activation of myogenic pathways and structural proteins in renal tissues, consistent with a pro-fibrotic phenotype. In contrast, SPK treatment reversed the expression of most genes, thereby restoring the gene profiles to those observed in control mice. Conclusions: MHV-1-induced long COVID is associated with persistent transcriptional reprogramming in the kidney, indicative of chronic inflammation, cytoskeletal dysregulation, and fibrogenesis. SPK demonstrates robust therapeutic potential by normalizing these molecular signatures and preventing long-term renal damage. These findings underscore the relevance of the MHV-1 model and support further investigation of SPK as a candidate therapy for COVID-19-associated renal sequelae.

The COVID-19 pandemic has been one of the most impactful events in our lifetime, caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Multiple SARS-CoV-2 variants were reported globally, and a wide range of symptoms existed. Individuals who contract COVID-19 continue to suffer for a long time, known as long COVID or post-acute sequelae of COVID-19 (PASC). While COVID-19 vaccines were widely deployed, both unvaccinated and vaccinated individuals experienced long-term complications. To date, there are no treatments to eradicate long COVID. We recently conceived a new approach to treat COVID in which a 15-amino-acid synthetic peptide (SPIKENET, SPK) is targeted to the ACE2 receptor binding domain of SARS-CoV-2, which prevents the virus from attaching to the host. We also found that SPK precludes the binding of spike glycoproteins with the receptor carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) of a coronavirus, murine hepatitis virus-1 (MHV-1), and with all SARS-CoV-2 variants. Further, SPK reversed the development of severe inflammation, oxidative stress, tissue edema, and animal death post-MHV-1 infection in mice. SPK also protects against multiple organ damage in acute and long-term post-MHV-1 infection. Our findings collectively suggest a potential therapeutic benefit of SPK for treating COVID-19.

The COVID-19 pandemic, which emerged in early 2020, has had a profound and lasting impact on global health, resulting in over 7.0 million deaths and persistent challenges. In addition to acute concerns, there is growing attention being given to the long COVID health consequences for survivors of COVID-19 with documented cases of cardiovascular abnormalities, liver disturbances, lung complications, kidney issues, and noticeable cognitive deficits. Recent studies have investigated the physiological changes in various organs following prolonged exposure to murine hepatitis virus-1 (MHV-1), a coronavirus, in mouse models. One significant finding relates to the effects on the gastrointestinal tract, an area previously understudied regarding the long-lasting effects of COVID-19. This research sheds light on important observations in the intestines during both the acute and the prolonged phases following MHV-1 infection, which parallel specific changes seen in humans after exposure to SARS-CoV-2. Our study investigates the histopathological alterations in the small intestine following MHV-1 infection in murine models, revealing significant changes reminiscent of inflammatory bowel disease (IBD), celiac disease. Notable findings include mucosal inflammation, lymphoid hyperplasia, goblet cell hyperplasia, and immune cell infiltration, mirroring pathological features observed in IBD. Additionally, MHV-1 infection induces villous atrophy, altered epithelial integrity, and inflammatory responses akin to celiac disease and IBD. SPIKENET (SPK) treatment effectively mitigates intestinal damage caused by MHV-1 infection, restoring tissue architecture and ameliorating inflammatory responses. Furthermore, investigation into long COVID reveals intricate inflammatory profiles, highlighting the potential of SPK to modulate intestinal responses and restore tissue homeostasis. Understanding these histopathological alterations provides valuable insights into the pathogenesis of COVID-induced gastrointestinal complications and informs the development of targeted therapeutic strategies.

Hepatic encephalopathy (HE) is a neurological condition linked to liver failure. Acute HE (Type A) occurs with acute liver failure, while chronic HE (Type C) is tied to cirrhosis and portal hypertension. HE treatments lag due to gaps in understanding its development by gender and age. We studied how sex and age impact HE and its severity with combined liver toxins. Our findings indicate that drug-induced (thioacetamide, TAA) brain edema was more severe in aged males than in young males or young/aged female rats. However, adding alcohol (ethanol, EtOH) worsens TAA’s brain edema in both young and aged females, with females experiencing a more severe effect than males. These patterns also apply to Type A HE induced by azoxymethane (AZO) in mice. Similarly, TAA-induced behavioral deficits in Type C HE were milder in young and aged females than in males. Conversely, EtOH and TAA in young/aged males led to severe brain edema and fatality without noticeable behavioral changes. TAA metabolism was slower in aged males than in young or middle-aged rats. When TAA-treated aged male rats received EtOH, there was a slow and sustained plasma level of thioacetamide sulfoxide (TASO). This suggests that with EtOH, TAA-induced HE is more severe in aged males. TAA metabolism was similar in young, middle-aged, and aged female rats. However, with EtOH, young and aged females experience more severe drug-induced HE as compared to middle-aged adult rats. These findings strongly suggest that gender and age play a role in the severity of HE development and that the presence of one or more liver toxins may aggravate the severity of the disease progression.

SignificancePreterm birth is defined as a birth before 37 weeks of gestation and is one of the leading contributors to infant mortality rates globally. Premature birth can lead to life-long developmental impairment for the child. Unfortunately, there is a significant lack of tools to diagnose preterm birth risk, which limits patient care and the development of new therapies.AimTo develop a speculum-free, portable preterm imaging system (PPRIM) for cervical imaging; testing of the PPRIM system to resolve polarization properties of birefringent samples; and testing of the PPRIM under an IRB on healthy, non-pregnant volunteers for visualization and polarization analysis of cervical images.ApproachThe PPRIM can perform 4×3 Mueller-matrix imaging to characterize the remodeling of the uterine cervix during pregnancy. The PPRIM is built with a polarized imaging probe and a flexible insertable sheath made with a compatible flexible rubber-like material to maximize comfort and ease of use.ResultsThe PPRIM device is developed to meet specific design specifications as a speculum-free, portable, and comfortable imaging system with polarized imaging capabilities. This system comprises a main imaging component and a flexible silicone inserter. The inserter is designed to maximize comfort and usability for the patient. The PPRIM shows high-resolution imaging capabilities at the 20 mm working distance and 25 mm circular field of view. The PPRIM demonstrates the ability to resolve birefringent sample orientation and full field capture of a healthy, non-pregnant cervix.ConclusionThe development of the PPRIM aims to improve access to the standard of care for women’s reproductive health using polarized Mueller-matrix imaging of the cervix and reduce infant and maternal mortality rates and better quality of life.

The coronavirus disease-19 (COVID-19) pandemic, declared in early 2020, has left an indelible mark on global health, with over 7.0 million deaths and persistent challenges. While the pharmaceutical industry raced to develop vaccines, the emergence of mutant severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) strains continues to pose a significant threat. Beyond the immediate concerns, the long-term health repercussions of COVID-19 survivors are garnering attention, particularly due to documented cases of cardiovascular issues, liver dysfunction, pulmonary complications, kidney impairments, and notable neurocognitive deficits. Recent studies have delved into the pathophysiological changes in various organs following post-acute infection with murine hepatitis virus-1 (MHV-1), a coronavirus, in mice. One aspect that stands out is the impact on the skin, a previously underexplored facet of long-term COVID-19 effects. The research reveals significant cutaneous findings during both the acute and long-term phases post-MHV-1 infection, mirroring certain alterations observed in humans post-SARS-CoV-2 infection. In the acute stages, mice exhibited destruction of the epidermal layer, increased hair follicles, extensive collagen deposition in the dermal layer, and hyperplasticity of sebaceous glands. Moreover, the thinning of the panniculus carnosus and adventitial layer was noted, consistent with human studies. A long-term investigation revealed the absence of hair follicles, destruction of adipose tissues, and further damage to the epidermal layer. Remarkably, treatment with a synthetic peptide, SPIKENET (SPK), designed to prevent Spike glycoprotein-1 binding with host receptors and elicit a potent anti-inflammatory response, showed protection against MHV-1 infection. Precisely, SPK treatment restored hair follicle loss in MHV-1 infection, re-architected the epidermal and dermal layers, and successfully overhauled fatty tissue destruction. These promising findings underscore the potential of SPK as a therapeutic intervention to prevent long-term skin alterations initiated by SARS-CoV-2, providing a glimmer of hope in the battle against the lingering effects of the pandemic.