Age-Stratified Infection Probabilities Combined With a Quarantine-Modified Model for COVID-19 Needs Assessments: Model Development Study.

BACKGROUND: Classic compartmental models such as the susceptible-exposed-infectious-removed (SEIR) model all have the weakness of assuming a homogenous population, where everyone has an equal chance of getting infected and dying. Since it was identified in Hubei, China, in December 2019, COVID-19 has rapidly spread around the world and been declared a pandemic. Based on data from Hubei, infection and death distributions vary with age. To control the spread of the disease, various preventive and control measures such as community quarantine and social distancing have been widely used. OBJECTIVE: Our aim is to develop a model where age is a factor, considering the study area's age stratification. Additionally, we want to account for the effects of quarantine on the SEIR model. METHODS: We use the age-stratified COVID-19 infection and death distributions from Hubei, China (more than 44,672 infections as of February 11, 2020) as an estimate or proxy for a study area's infection and mortality probabilities for each age group. We then apply these probabilities to the actual age-stratified population of Quezon City, Philippines, to predict infectious individuals and deaths at peak. Testing with different countries shows the predicted number of infectious individuals skewing with the country's median age and age stratification, as expected. We added a Q parameter to the SEIR model to include the effects of quarantine (Q-SEIR). RESULTS: The projections from the age-stratified probabilities give much lower predicted incidences of infection than the Q-SEIR model. As expected, quarantine tends to delay the peaks for both the exposed and infectious groups, and to "flatten" the curve or lower the predicted values for each compartment. These two estimates were used as a range to inform the local government's planning and response to the COVID-19 threat. CONCLUSIONS: Age stratification combined with a quarantine-modified model has good qualitative agreement with observations on infections and death rates. That younger populations will have lower death rates due to COVID-19 is a fair expectation for a disease where most fatalities are among older adults.

Increased association of the ERG oncoprotein expression in advanced stages of prostate cancer in Filipinos.

BACKGROUND: Filipinos with prostate cancer (CaP) are at increased risk of harboring advanced stages and lower survival rates compared to other Asians. This study aims to investigate prevalence of ETS-related gene (ERG) oncoprotein overexpression in Filipinos as surrogate of TMPRSS2-ERG gene fusions, using a highly specific monoclonal antibody (ERG-MAb), and conduct the first attempt to study the role of genetic alterations in the aggressive tumor biologic behaviour of CaP among Filipinos. METHODS: This case-matched, case-control retrospective study evaluated ERG expression in Filipino patients diagnosed with CaP and its effect on stage and Gleason grade of their disease. Men who underwent radical prostatectomy for organ-confined disease at the University of the Philippines-Philippine General Hospital (UP-PGH) comprised the organ-confined cohort. Age-matched adults who had trans-rectal ultrasound-guided prostate (TRUSP) biopsy or trans-urethral resection of the prostate (TURP) with bilateral orchiectomy for T4 or stage IV CaP composed the advanced disease cohort. RESULTS: Overall ERG expression frequency of 23.08% (N = 104) was demonstrated, with a higher rate observed in the advanced disease cohort (32.69%) compared to the organ-confined group (13.46%). Furthermore, ERG overexpression was only detected among intermediate and high-risk tumors. A high-specificity (98.08%) of the ERG-MAb for malignant prostatic cells was likewise demonstrated. CONCLUSIONS: In contrast to higher ERG frequency in Western countries, it is much lower in Filipino CaP, which is similar to lower rates noted from other Asian countries. The 98.08% specificity of ERG oncoprotein for prostate tumor cells combined with its increased association in advanced disease, suggests for prognostic potential of ERG that may aid clinicians in treatment decisions for Filipino CaP patients.

The COVID-19 pandemic's effects on mental and psychosocial health in the Philippines: A scoping review.

Low- and middle-income countries (LMICs) remained at elevated risk for the effects of the COVID-19 pandemic because of persistent stressors to their health systems. Simultaneously facing high infection rates, strict containment measures and natural disasters, the Philippines provides important grounds for health research in LMICs. This review examined how the COVID-19 pandemic affected mental and psychosocial health in the Philippines. This scoping review included literature in English from 2020 to mid-2022 from PubMed, PsycInfo and SCOPUS, and used the PRISMA-ScR and PCC-question model. Two independent reviewers conducted blind article screening and data extraction using COVIDENCE software, followed by consensus building, data charting and analyses. This work identified 405 publications across PubMed (N = 56), PsycInfo (N = 106) and SCOPUS (N = 243), of which 76 articles addressed the Philippines. Article types included 54 research articles, 10 opinion pieces, 4 literature reviews, 6 letters to journals, 1 study protocol and 1 other report. These findings focused primarily on health professionals (N = 23) and educators/learners (N = 22) and reported mostly on moderate-to-severe clinical outcomes such as fear, depression, anxiety or stress. Coping behaviors, like resiliency and other ways of adapting to the pandemic, including religious, spiritual and community-oriented approaches highlighted experiences with stringent infection prevention and control measures to contain COVID-19 in the Philippines. The COVID-19 pandemic brought severe challenges to mental and psychosocial health in the Philippines. The literature focused mostly on healthcare workers and educators/learners, and moderate-to-severe mental health outcomes in these groups. There is a need to expand studies to other sociodemographic groups and communities across the Philippines. Future work stands to benefit from more in-depth qualitative, mixed methods, longitudinal and representative quantitative research in LMICs following this pandemic. Literature reviews remain important to synthesize post-pandemic experiences by providing context for future studies and health practice in the Philippines and other LMICs.

Novel in vitro and in vivo models and potential new therapeutics to break the vicious cycle of Cryptosporidium infection and malnutrition.

BACKGROUND: Although several animal models of cryptosporidiosis have been reported, most involve genetically or pharmacologically immune-suppressed hosts. METHODS: We report challenge with excysted (in vitro and in vivo) and unexcysted (in vivo) Cryptosporidium parvum oocysts in human colonic adenocarcinoma (HCT-8) cells and weaned nourished and malnourished C57BL/6 mice, following outcomes of growth rate, stool shedding, and tissue burden. We tested treatment with an oligodeoxynucleotide containing unmethylated CpG motif (CpG-ODN) and alanyl-glutamine in vivo and in vitro. RESULTS: C. parvum-challenged mice showed prolonged weight loss (>10% over 4 days), robust stool shedding (>3 logs/d over 7 days), and epithelial infection in the ileum, cecum, and colon. Of 2 potential therapeutic compounds evaluated in the model, CpG-ODN reduced body weight loss (to <6% on days 3-7 after challenge), reduced shedding of organisms (by 25% on days 1 and 3 after challenge), and decreased the burden of parasites in the ileum. Alanyl-glutamine showed similar benefits. In vitro findings suggested that effects on the epithelial component of the mucosa probably likely responsible for beneficial effects seen in vivo. CONCLUSIONS: Weaned mice provide a convenient and reproducible model of cryptosporidial disease, including its vicious cycle with body weight loss and heavier infection with malnutrition, and this model may be useful in exploring innovative therapeutic solutions for this challenging infectious disease.

Cryptosporidium infection causes undernutrition and, conversely, weanling undernutrition intensifies infection.

Cryptosporidium parvum is a leading pathogen in children in developing countries. To investigate whether early postnatal malnutrition leads to heavier C. parvum infections, we assessed intestinal adaptation and parasite load in suckling mice during the first 2 wk of life, analogous to the first postnatal yr in humans. Undernutrition was induced by daily C57BL6J pup separation from lactating dams. Half of the pups were separated daily, for 4 hr on day 4, 8 hr on day 5, and for 12 hr from day 6 until day 14. On day 6, each pup received an oral inoculum of 10(5) to 10(7) parasites in 10-25 microl of PBS. Littermate controls received PBS alone. Stools were assessed from days 8, 11, and 14 for oocyst counts. Mice were killed on day 14, 8 days postinoculation, at the peak of the infection. Ileal and colon segments were obtained for histology, real-time and reverse transcriptase PCR, and immunoassays. Villus and crypt lengths and cross-sectional areas were also measured. Undernourished and nourished mice infected with excysted 10(6) or 10(7) oocysts exhibited the poorest growth outcomes compared with their uninfected controls. Nourished 10(6)-infected mice had comparable weight decrements to uninfected undernourished mice. Body weight and villi were additively affected by malnutrition and cryptosporidiosis. Hyperplastic crypts and heavier inflammatory responses were found in the ilea of infected malnourished mice. Undernourished infected mice exhibited greater oocyst shedding, TNF-alpha and IFN-gamma intestinal levels, and mRNA expression compared to nourished mice infected with either 10(5) or 10(6) oocysts. Taken together, these findings show that Cryptosporidium infection can cause undernutrition and, conversely, that weanling undernutrition intensifies infection and mucosal damage.

Detection and quantification of Cryptosporidium in HCT-8 cells and human fecal specimens using real-time polymerase chain reaction.

Cryptosporidium is a significant cause of diarrheal illness worldwide, especially among children and immunocompromised patients. Currently used diagnostic techniques are time-consuming, require skilled technicians, and are not useful for quantification of oocysts in fecal and environmental samples. In this study, we examined the use of a real-time polymerase chain reaction (PCR) assay for detecting and quantifying Cryptosporidium parvum in three distinct and progressively more complex matrices: phosphate-buffered saline (PBS), HCT-8 cells (human ileocecal carcinoma), and human fecal specimens. A reliable standard curve was generated using the PBS samples spiked with pure oocysts, and oocyst starting quantities were calculated for the infected HCT-8 cell and spiked fecal samples. The assay detected Cryptosporidium in samples infected/spiked with > or =10(3) oocysts/sample and detected both C. hominis and C. parvum in clinical specimens. This assay is useful in a variety of samples in the research laboratory and will likely prove to be a useful tool in the clinical laboratory.

High anti-Cryptosporidium parvum IgG seroprevalence in HIV-infected adults in Limpopo, South Africa.

A seroepidemiological study was performed to determine the seroprevalence of Cryptosporidium in human immunodeficiency virus (HIV)-infected adults and local university students in the Limpopo Province, South Africa. Using a custom anti-C. parvum immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA), the seroprevalence of Cryptosporidium was found to be significantly higher (75.3%; 146 of 193) in HIV-infected individuals compared with student volunteers (32.8%; 19 of 58) (P < 0.001). A more recent diagnosis of HIV was associated with anti-C. parvum IgG seropositivity, as was lower weight among HIV-infected women. This is the first seroepidemiologic study of Cryptosporidium in rural South Africa, and it shows high endemicity among the HIV-infected population. In addition to raising the possibility of significant Cryptosporidium-related morbidities, this finding reveals that in Limpopo and perhaps in other low-income, rural populations, interrupting waterborne pathogen transmission will require strategies effective against environmentally hardy parasites such as Cryptosporidium.

Clostridium difficile and Entamoeba histolytica infections in patients with colitis in the Philippines.

Amoebiasis is a common cause of non-specific colitis in the Philippines. The prevalence of Clostridium difficile infection with colitis is unknown. Empiric use of metronidazole for colitis treatment is widely practiced. We investigated the association of C. difficile or Entamoeba histolytica infection with endoscopically/histopathologically proven colitis among adults in the Philippines. Two hundred and ten patients undergoing colonoscopy were enrolled. Demographic and clinical data were reviewed. Stool specimens were assayed for C. difficile and E. histolytica by ELISA. Microscopy was performed. The mean age of the patients was 53 y (range: 19-88 y) and 53% were male. Colitis was diagnosed in 39 of 205 patients. Clostridium difficile, E. histolytica and parasites were seen in 17 (43.6%), 10 (25.6%) and 11 (28.2%), respectively, of patients with colitis compared with 36 (21.7%; p=0.005), 13 (7.8%; p=0.001) and 56 (33.7%; p=0.51), respectively, of those without colitis. Diarrhoea and antibiotic intake history were significantly more common among patients with colitis than those without (43.6% and 20.5% vs 18.1% and 5.4%; p=0.001 and p=0.006, respectively). The mean duration of diarrhoea was 2.53 d shorter among patients with colitis. The most frequent antibiotics taken were fluoroquinolones and metronidazole (50% and 40% of antibiotic courses, respectively, in patients with colitis). This study suggests that C. difficile infection is common and might be overlooked in settings where amoebiasis and intestinal parasitism are endemic.

Arginine decreases Cryptosporidium parvum infection in undernourished suckling mice involving nitric oxide synthase and arginase.

OBJECTIVE: This study investigated the role of L-arginine supplementation to undernourished and Cryptosporidium parvum-infected suckling mice. METHODS: The following regimens were initiated on the fourth day of life and injected subcutaneously daily. The C. parvum-infected controls received L-arginine (200 mmol/L) or phosphate buffered saline. The L-arginine-treated mice were grouped to receive NG-nitro-arginine methyl ester (L-NAME) (20 mmol/L) or phosphate buffered saline. The infected mice received orally 10(6) excysted C. parvum oocysts on day 6 and were euthanized on day 14 at the infection peak. RESULTS: L-arginine improved weight gain compared with the untreated infected controls. L-NAME profoundly impaired body weight gain compared with all other groups. Cryptosporidiosis was associated with ileal crypt hyperplasia, villus blunting, and inflammation. L-arginine improved mucosal histology after the infection. L-NAME abrogated these arginine-induced improvements. The infected control mice showed an intense arginase expression, which was even greater with L-NAME. L-arginine decreased the parasite burden, an effect that was reversed by L-NAME. Cryptosporidium parvum infection increased urine NO(3)(-)/NO(2)(-) concentrations compared with the uninfected controls, which was increased by L-arginine supplementation, an effect that was also reversed by L-NAME. CONCLUSION: These findings show a protective role of L-arginine during C. parvum infection in undernourished mice, with involvement of arginase I and nitric oxide synthase enzymatic actions.

Cryptosporidium-malnutrition interactions: mucosal disruption, cytokines, and TLR signaling in a weaned murine model.

Cryptosporidiosis is a leading cause of persistent diarrhea in children in impoverished and developing countries and has both a short- and long-term impact on the growth and development of affected children. An animal model of cryptosporidial infection that mirrors closely the complex interaction between nutritional status and infection in children, particularly in vulnerable settings such as post-weaning and malnourishment, is needed to permit exploration of the pathogenic mechanisms involved. Weaned C57BL/6 mice received a protein-deficient (2%) diet for 3-12 days, then were infected with 5 × 10(7) excysted C. parvum oocyts, and followed for rate of growth, parasite stool shedding, and intestinal invasion/morphometry. Mice had about 20% reduction in weight gain over 12 days of malnutrition and an additional 20% weight loss after C. parvum challenge. Further, a significantly higher fecal C. parvum shedding was detected in malnourished infected mice compared to the nourished infected mice. Also, higher oocyst counts were found in ileum and colon tissue samples from malnourished infected mice, as well as a significant reduction in the villous height-crypt depth ratio in the ileum. Tissue Th1 cytokine concentrations in the ileum were significantly diminished by malnutrition and infection. mRNA for toll-like receptors 2 and 4 were diminished in malnourished infected mice. Treatment with nitazoxanide did not prevent weight loss or parasite stool shedding. These findings indicate that, in the weaned animal, malnutrition intensifies cryptosporidial infection, while cryptosporidial infection further impairs normal growth. Depressed TLR2 and 4 signaling and Th1 cytokine response may be important in the mechanisms underlying the vicious cycle of malnutrition and enteric infection.

Diarrhea, clostridium difficile, and intestinal inflammation in residents of a long-term care facility.

INTRODUCTION: Long-term care facilities (LTCF) residents have been estimated to have the highest incidence of diarrheal illness among adults living in the developed world. This study describes undiagnosed diarrhea, intestinal inflammation, and Clostridium difficile colonization in a LTC population and explores whether these are associated with functional decline, as defined by weight loss or a change in cognitive or ADL status. METHODS: An observational study of a convenience sampling of residents in a 180-bed LTCF was obtained; evaluation of stool and medical records was done. Stool specimens were evaluated for consistency, gross blood, inflammation (via quantitative fecal lactoferrin, IBD-SCAN), and C difficile (via PCR for gdh). SPSS and STATA were used and significance was set at P < .05. RESULTS: There were 46 stools collected; 13 of the subjects were male, 28 were older than 65 years, and 35 were prescribed 5 to 15 medications. Twenty-six of the 46 stools collected had elevated quantitative fecal lactoferrin levels. Although only 5 subjects were reported to have diarrhea (4 with elevated lactoferrin), 28 stool specimens were observed to be liquid or semi-solid (19 with elevated lactoferrin), and these liquid/ semisolid stools were significantly correlated with lactoferrin positivity (P = .017). In analysis of functional status, there was no statistically significant association between change in ADL (n = 17) or cognitive status (n = 5) and elevated lactoferrin. However, all 3 subjects who had significant weight loss had elevated lactoferrin, although the mean fecal lactoferrin was not statistically different from those without weight loss. Of the 2 samples with C difficile, both were liquid and, when compared with all other liquid stools (n = 22), the mean lactoferrin was statistically higher (134.1 versus 28.8 microg/mL, P = .008). These 2 subjects had neither weight loss nor change in cognitive status, but 1 had a change in ADL status. DISCUSSION AND CONCLUSIONS: Diarrhea in LTCF residents is underdiagnosed. Diarrhea and the presence of C difficile in the stool are associated with intestinal inflammation, as detected by fecal lactoferrin. With our small numbers, we were not able to identify a specific link; however, we were able to identify a correlation between weight loss and intestinal inflammation, but, with just 2 samples, not C difficile colonization. This relationship highlights the importance of larger studies to further examine the rate of diarrhea in LTCF; the effect of diarrhea and intestinal inflammation on weight loss; and the interaction of C difficile colonization with weight loss, malnutrition, and functional decline.

Detection of epithelial-cell injury, and quantification of infection, in the HCT-8 organoid model of cryptosporidiosis.

BACKGROUND: Intestinal cells grown in microgravity produce a three-dimensional tissue assembly, or "organoid," similar to the human intestinal mucosa, making it an ideal model for enteric infections such as cryptosporidiosis. METHODS: HCT-8 cells were grown in a reduced-gravity, low-shear, rotating-wall vessel (RWV) and were infected with Cryptosporidium parvum oocysts. Routine and electron microscopy (EM), immunolabeling with fluorescein-labeled Vicia villosa lectin and phycoerythrin-labeled monoclonal antibody to a 15-kD surface-membrane protein, and quantitative polymerase chain reaction (qPCR) using probes for 18s rRNA of C. parvum and HCT-8 cells were performed. RESULTS: The RWV allowed development of columnar epithelium-like structures. Higher magnification revealed well-developed brush borders at the apical side of the tissue. Incubation with C. parvum resulted in patchy disruption of the epithelium and, at the surface of several epithelial cells, in localized infection with the organism. EM revealed irregular stunting of microvilli, foci of indistinct tight junctions, and areas of loose paracellular spaces. qPCR showed a 1.85-log (i.e., 70-fold) progression of infection from 6 h to 48 h of incubation. CONCLUSION: The HCT-8 organoid displayed morphologic changes indicative of successful and quantifiable infection with C. parvum. The HCT-8 organoid-culture system may have application in interventional in vitro studies of cryptosporidiosis.