Hypoadrenocorticism (HA) in dogs requires lifelong substitution therapy with glucocorticoids and, in most cases, also mineralocorticoids. For glucocorticoid replacement, prednisolone is administered at a maintenance dose of approximately 0.1 mg/kg/day. Alternative drugs such as hydrocortisone, dexamethasone, or methylprednisolone are used less frequently.

Hypoadrenocorticism in cats is a rare endocrine disorder that should be considered in patients presenting with vague, intermittent, or waxing and waning clinical signs. Chronic gastrointestinal signs, poor growth, or episodes of hypovolemic shock should raise clinical suspicion. British Shorthair cats may be overrepresented among affected cats, and clinicians should be particularly alert when this breed presents with compatible signs. Definitive diagnosis requires an ACTH stimulation test, which should be performed prior to the initiation of any glucocorticoid treatment to avoid diagnostic interference. In clinically stable cats, baseline cortisol measurement may be used as an initial screening tool. However, as no feline-specific thresholds have been validated, the commonly used cut-off of >2 µg/dL (>55 nmol/L) to rule out hypoadrenocorticism should be interpreted with caution, as it may be less reliable than in dogs. Long-term management involves lifelong glucocorticoid replacement, typically using oral prednisolone. Cats generally require higher maintenance doses than dogs to maintain clinical stability. Mineralocorticoid deficiency is managed with subcutaneous administration of an extended-release desoxycorticosterone preparation (Zycortal®) given monthly at a starting dose of 2,2 mg/kg - substantially higher than canine doses. As an alternative, daily oral fludrocortisone acetate (Florinef® or Astonin®H) could be used at 0,01-0,02 mg/kg/day in selected cases. Regular monitoring of sodium and potassium levels is essential to adjust mineralocorticoid dosages and optimize treatment. Because affected cats cannot mount an adequate stress response, additional glucocorticoid supplementation («glucocorticoid boost») is required during stressful situations or concurrent illness. Owners must be educated about the signs of adrenal crisis, which can be more subtle and difficult to recognize in cats compared to dogs. Emphasis should be placed on consistent medication administration and routine veterinary monitoring to ensure long-term disease control and quality of life.

Low-dose oral formulations are often associated with poor dose accuracy (mass and content non-uniformity) and limited reproducibility. Three-dimensional printing (3DP), particularly Fused Deposition Modeling (FDM), has emerged as a promising strategy to overcome limitations of conventional pharmaceutical compounding for oral therapies. FDM enables precise, digitally controlled deposition for decentralized, on-demand manufacturing at the Point of Care (PoC), but its implementation is limited by the availability of suitable pharmaceutical filaments. Especially at sub-percent drug loadings, achieving homogeneous drug distribution at such low concentrations is a major challenge. Fludrocortisone Acetate (FCA), a micronized corticosteroid administered at microgram daily doses and commonly compounded in Belgium in the absence of authorized products, was selected as a model drug. This study aimed to develop homogeneous low-dose (0.5% w/w) FCA-loaded filaments by Hot-Melt Extrusion (HME) suitable for FDM printing. The effects of adding 1% of a flow regulator (Aerosil® 200) and screw configuration (one vs. two kneading zones) on extrusion process parameters and drug distribution were investigated. Aerosil® 200 improved powder flow and feeding stability, and filament homogeneity resulted from the combined effect of formulation and screw configuration. Optimized filaments showed suitable mechanical properties for FDM and enabled printing of Immediate-Release (IR) oral dosage forms containing 50, 100, and 200 µg FCA. Printed waffle-shaped units complied with European Pharmacopoeia requirements for mass and content uniformity, hardness, and friability. Dissolution confirmed an IR profile, with > 80% FCA released within 45 min. This work demonstrates the feasibility of manufacturing pharmaceutically compliant low-dose dosage forms by combining HME and FDM, offering a reliable alternative to conventional compounding and advancing point-of-care personalized medicine.

Vasovagal syncope is a frequent problem affecting nearly half the population. The symptom burden is wide, ranging from once to hundreds of times. It reduces quality of life and causes injuries. Physicians, particularly front-line physicians such as family doctors, need a simple, practical approach.

Hypertension is one of the most common factors contributing to cardiac remodeling that links to changes in both heart and kidney functions. Several studies have reported the beneficial effects of Coenzyme Q10 (CoQ10) in reducing oxidative stress due to its antioxidant properties. However, the cardioprotective and nephroprotective roles of CoQ10 against the toxicity induced by fludrocortisone acetate (FCA) have yet to be investigated. Therefore, we evaluated the CoQ10 effects on FCA-induced cardiotoxicity and nephrotoxicity in a rat model (FCA-Salt). We analyzed various oxidative stress parameters and antioxidant enzyme activities using biochemical assays. In addition, we examined inflammatory and fibrosis via histological staining of heart and kidney tissues. CoQ10 significantly reduced lipid peroxidation product-malondialdehyde (MDA) while increased nitric oxide (NO) in the FCA-Salt rats. Antioxidant enzyme activities were also enhanced by CoQ10 in these rats. Histopathological examination detected tissue damage and fibrosis in the FCA-Salt group, which was significantly reduced by CoQ10. Furthermore, enzyme-linked immunosorbent assay (ELISA) indicated the activation of Transforming growth factor-β1/Tumor necrosis factor-α (TGF-β1/TNF-α) by CoQ10. Consequently, our study suggests that CoQ10 can effectively protect heart and kidney health against cardiotoxicity and nephrotoxicity in FCA-Salt hypertensive rats via TGF-β1/TNF-α pathway.

Uncontrolled hypertension is a primary contributor to tissue damage in multiple organs, including the heart and kidneys. In this study, we explored the protective roles of L-arginine in a mineralocorticoid-induced rat model of hypertension. To induce hypertension and subsequent organ damage, rats were nephrectomized unilaterally and fed with the mineralocorticoid fludrocortisone acetate and dietary salt (FCA-Salt). These rats were treated with L-arginine for 28 days, and subsequent tests were performed. Biochemical analysis revealed the increased level of inflammation and oxidative stress biomarkers in the plasma, heart, and kidney of the FCA-salt-treated rats. L-arginine treatment decreased the oxidative stress marker malondialdehyde (MDA) by 18 %, 22 %, and 18 % in the heart, kidney, and plasma, respectively. L-arginine also attenuated the advanced oxidative protein products (AOPP). The activity of superoxide dismutase (SOD) increased by 62 %, 45 %, and 16 % in the heart, kidney, and plasma, respectively, in the L-arginine-treated animals compared to the FCA-Salt group. Significant augmentation was also revealed for the nitric oxide (NO), catalase (CAT), and reduced glutathione (GSH). The plasma levels of the kidney function biomarkers uric acid and creatinine were significantly improved after L-arginine treatment. Furthermore, L-arginine remarkably reduced the elevated plasma cytokines IL-1β, IL-17A, TNF-α, and TGF-β1 in FCA-salt-induced hypertensive rats. Histopathological data showed a reduction in fibrosis and tissue damage by L-arginine in the FCA-salt-treated group. We propose that L-arginine could be an effective agent in preventing cardiac and renal dysfunction in hypertensive rats.

Hypertensive rats serve as a good experimental model for studying the pathophysiology of cardiac hypertrophy and remodeling leading to heart failure. In this study, we aimed to analyze the effect of astaxanthin and possible mechanisms involved in alleviating oxidative stress, fibrosis and inflammation that triggers cardiac remodeling using male uninephrectomized Long Evans rats. Cardiac hypertrophy and hypertension were induced in rats termed as 'FCA-Salt rats' by an oral administration of fludrocortisone acetate (FCA) and 1 % NaCl in drinking water. Biochemical assays showed that FCA-Salt rats exhibited an upregulation of oxidative stress markers AOPP, MDA and downregulation of NO in heart and kidney, which was reversed by astaxanthin treatment. Astaxanthin further regularized the reduced activities of antioxidant enzymes GSH, SOD and CAT in these tissues. ELISA revealed that astaxanthin significantly reduced the inflammatory response by reducing the elevated levels of IL-1β, IL-17a, and TNF-α and pro-fibrotic marker TGF-β1 in plasma. Real-time qPCR depicted an upregulation of TNF-α, IL-1β, IL-6, IL-17A as well as signaling molecules TGF-β1, Smad2 and Smad3 in heart of FCA-Salt rats, which was reduced significantly by astaxanthin. Sirius red staining showed that the cardiac and renal fibrosis was significantly improved by astaxanthin treatment. Together, our results suggest that astaxanthin treatment is beneficial in protecting cardio-renal damage in hypertension through TGF-β/Smad signaling pathway, hence, this molecule may be considered for the maintenance of cardio-renal health.

Engineered bacterial magnetic nanoparticles (BMPs) fused with protein A (BMP-PA) can bind antibodies, creating immunomagnetic beads that offer an attractive tool for targets screening. In the study, BMP-PA-IgG was formed by attaching broad-spectrum monoclonal antibodies against glucocorticoids (GCs) to BMP-PA. Immunomagnetic assay was developed for analysis of GCs, using the BMP-PA-IgG and hydrocortisone-horseradish peroxidase. The developed assay exhibited broad specificity for GCs, including hydrocortisone (HCS), betamethasone (BMS), dexamethasone (DMS), prednisolone (PNS), beclomethasone (BCMS), cortisone (CS), 6-α-methylprednisone (6-α-MPNS), and fludrocortisone acetate (HFCS), with half inhibitory concentrations (IC50) ranging from 0.88 to 6.57 ng/mL. The proposed assay showed average recoveries of HCS and DMS ranging from 75.6% to 105.2% in chicken and pork samples, which were correlated well with those obtained by LC-MS/MS. This study indicated that the integration of engineered immunomagnetic beads into immunoassay systems offer possibilities for the sensitive and selective detection of GCs.

Immunomagnetic beads provide novel tools for high-throughput immunoassay techniques. In this study, protein G (PG) was immobilized onto bacterial magentic particles (BMPs) using an additional cysteine residue at the C-terminus. A broad-spectrum monoclonal antibody against glucocorticoids (GCs) was attached to BMPs through PG-Fc interaction, generating BMP-PG-mIgG immunomagentic beads. A sensitive one-step immunoassay was developed for GCs based on combination of BMP-PG-mIgG and dexamethasone-horseradish peroxidase tracer (DMS-HRP). The developed assay exhibited half inhibitory concentrations (IC50) for dexamethasone (DMS), betamethasone (BMS), prednisolone (PNS), hydrocortisone (HCS), beclomethasone (BCMS), cortisone (CS), 6-α-methylprednisone (6-α-MPNS), fludrocortisone acetate (HFCS) of 0.98, 1.49, 2.42, 9.29, 1.63, 6.13, 7.3, and 4.89 ng/mL, respectively. The method showed recoveries ranging rates from 86.5 % to 117 % with a coefficient of variation less than 12.3 % in milk sample, which showed a good correlation with LC-MS/MS. Thus, the proposed assay offers a rapid and broad-spectrum screening tool for simultaneous detection of GCs in milk.

A 12-year-old neutered male Chihuahua dog was diagnosed with pituitary-dependent hypercortisolism and treated with trilostane. Eighty-nine days later, the dog showed lethargy accompanied by hyponatraemia and hyperkalaemia. Hypoadrenocorticism due to trilostane was suspected, but the result of the adrenocorticotropic hormone stimulation test was not conclusive. Contrast-enhanced ultrasound showed loss of adrenocortical blood flow in both adrenal glands, indicating adrenocortical hypoperfusion and isolated hypoadrenocorticism. Treatment with fludrocortisone acetate improved the condition and electrolyte abnormalities. Thirteen months later, the dog showed alopecia, and an adrenocorticotropic hormone stimulation test revealed increased cortisol concentration, indicating hypercortisolism recurrence. The dog died due to progressive deterioration 22 months after the initial presentation. Post-mortem examination revealed focally extensive necrosis with marked calcification in the parenchyma of the adrenal glands and regeneration of the cells in the zona fasciculata with severe fibrosis. Adrenocortical hypoperfusion detected by contrast-enhanced ultrasound can support the diagnosis of adrenal necrosis and hypoadrenocorticism.

Congenital adrenal hyperplasia (CAH) is a rare condition usually referred to as a group of genetic disorders resulting due to a deficiency of steroid enzymes required by adrenal glands to produce cortisol and mineralocorticoid hormones. It has an autosomal recessive mode of inheritance and is further categorized into two types-Classic and Non-Classic. Non-Classic CAH is a more common milder form that presents late after puberty. Classic CAH, although more severe, is rare and detected at birth and is associated with the life-threatening adrenal crisis in both sexes and virilization of the external genitalia in females (46, XX) patients, whereas in males, no overt abnormality of the external genitalia is present. We present a case of a four-month-old male child with the classic form of CAH who was brought with complaints of loose stools, projectile non bilious vomiting, decreased urine output, and failure to feed for 3 days. The child had a clinical presentation of salt wasting with hypoglycemia and hyperpigmentation of his genitalia. The USG findings revealed increased anteroposterior diameter of renal pelvis indicative of a growth in the suprarenal area. 17-hydroxyprogesterone (17-OHP) was found to be elevated confirming the diagnosis. He was treated with hydrocortisone with gradual improvement in his glucose and electrolytes. The patient was discharged home on replacement therapy consisting of oral prednisolone and fludrocortisone acetate and followed up as outpatient with significant improvement in the clinical findings. The fact that the child was not screened for CAH at birth led to the critical consequences of the disease in this case. To prevent life-threatening adrenal crisis and help perform appropriate sex assignments for affected female patients, newborn screening (NBS) programs for the classical form of CAH should be made mandatory even in low- and middle-income countries.

Fludrocortisone acetate is a drug used to treat adrenal insufficiencies which can be prescribed to hospitalized or ambulatory pediatric patients at dosages not commercially available. For these patients, 10-µg fludrocortisone capsules are currently compounded from a pre-compounded titrated powder (powder triturate). Fludrocortisone stability studies were carried out to ensure a valid beyond-use date. First, a stability-indicating fludrocortisone acetate dosing method was validated. Then fludrocortisone acetate 10-µg capsules and 1% fludrocortisone acetate titrated powders (powder triturates) were realized. Finally, stability studies were performed. The fludrocortisone acetate titrated powders (powder triturates) were stable for one year at controlled ambient temperature and protected from light, whereas 10-µg fludrocortisone acetate capsules were stable for six months. One year after, even if the fludrocortisone content remained conformed, an increase in product degradation was noted. Our work allowed us to determine a six-month beyond-use date for fludrocortisone acetate titrated powder (powder triturate) with the three most commonly used excipients for capsule compounding. We also confirmed the sixmonth theoretical stability for capsules.