Background: Allovahlkampfia spelaea (A. spelaea) is a free-living amoeba that has recently been recognized to
cause Acanthamoeba-like keratitis, the treatment of which is complex. The pathogenic potential of Allovahlkampfia
spp. remains unexplored. This study characterized A. spelaea through ultrastructural morphological
analysis and investigated the pathogenic potential of the A. spelaea strain KS1, which was isolated from a patient
with keratitis, in a murine model, with a focus on neuro-pulmonary infections. Additionally, this study assessed
the therapeutic effectiveness of ellagic acid (EA) against tissue damage caused by amoebic infections.
Methods: Immunosuppressed male Wister rats were intranasally inoculated with A. spelaea trophozoites (1 × 106/
ml) and divided into control, infected untreated, and infected treated (50 mg/kg EA daily) groups. Histopathological
and ultrastructural analyses of brain and lung tissues were conducted by scanning and transmission
electron microscopy. Additionally, the therapeutic effects of EA were assessed via comparative tissue pathology.
Results: A. spelaea infection induced A. spelaea-induced neural lesions resembling granulomatous amoebic encephalitis
(GAE) in the brain, which was characterized by gliosis, vasculitis, and necrosis, in addition to severe
pulmonary damage, including suppurative bronchopneumonia and abscesses. Trophozoites presented with
pseudopodia, acanthopodia, and amoebostomes, whereas cysts presented with double-layered walls. EA-treated
rats presented nearly normal brain and lung histology, with reduced inflammation and gliosis, highlighting the
anti-inflammatory and antioxidant properties of EA.
Conclusion: This study highlights the neurotropic and pulmonary pathogenicity of A. spelaea, with ultrastructures
parallel to those of Vahlkampfia spp. and Acanthamoeba spp. Ellagic acid significantly reduces infection-induced
damage, underscoring its potential as a therapeutic agent for infections caused by free-living amoebae.

Abstract
Objective This study’s aim was: (1) introduce the digital drying oven as a reproducible, controllable, and accurate
heating device for burn model creation. (2) Define the heating temperature appropriate for developing consistent
second and third-degree burn injuries in rats.
Results Burns appeared deeper with more distinct borders in groups (B) and (C) than in group (A). The stainless-steel
rod at 100 ºC created burn injuries of the second degree, evidenced by the sloughing of the epidermis and necrosis
in the epithelium and upper part of the dermis. Heating at 150 and 200 ºC created third-degree burn injuries, where
necrosis involved the epidermis and dermis and extended to the subcutaneous fat and muscles. The depth of the
burn wound in the group (B) (371.2 ± 41.3 μm) and (C) (385.2 ± 38.0 μm) was significantly deeper compared with the
group (A) (178 ± 46.6 μm) (P < 0.001). The digital drying oven is a reliable, reproducible, and controllable heating device
for creating burn models. The stainless-steel rod (63 g and 8 mm) heated at 100 and 150 ºC with a contact time of 30 s
is adequate for creating consistent second and third-degree burn injuries in rats, respectively.

Background

T2DM is a chronic disorder with progressive neuromuscular alterations. L-arginine (ARG) is the most common semi-essential amino acid having several metabolic functions.

Aim

to investigate the impact of L-arginine in combating diabetic-induced neuromyopathy and its possible mechanisms.

Materials & Methods

24 rats were divided into CON, CON+ARG, DC, DC+ARG. Behavioral tests, Body weight (BW), fasting blood glucose (FBG), insulin, total antioxidant capacity (TAC), malondialdehyde (MDA), plasminogen activator inhibitor-1 (PAI-1), and irisin were done. Creatine kinase-MM (CK-MM), interleukin 4 (IL-4), interleukin 6 (IL-6), TAC, MDA, expression of microRNA-29a mRNA & light chain 3 protein were determined in muscle. Histological and NF-κβ immunohistochemical expression in muscle and nerve were assessed.

Results

ARG supplementation to diabetic rats improved altered behavior, significantly increased BW, insulin, TAC, irisin and Il-4, decreased levels of glucose, microRNA-29a, NF-κβ and LC3 expression, PAI-1, CK-MM and restored the normal histological appearance.

Conclusions

ARG supplementation potently alleviated diabetic-induced neuromuscular alterations.

This study evaluated the application of chitosan/polyvinyl alcohol/graphene oxide/nano titanium oxide (CS/PVA/GO/nano TiO₂) hydrogels for bone defect reconstruction in dogs. Dogs were subjected to mid-diaphyseal circular bone defects (0.8 cm²) in the radius bones. Bone defects were implanted with the hydrogel in the treated group (n = 9), while the control group were subjected to spontaneous healing (n = 9). Dogs were subjected to clinical, radiographic, and scanning electron microscopy (SEM) evaluations at 15-, 30-, and 45-days post-surgery. Dogs in the treated group recorded no lameness by the end of the third week post-surgery, while dogs in the untreated group still exhibited lameness of grade 1. There was a significant decrease (p < 0.05) in the cortical defect (mm) of the treated group (5.46 ± 0.17 and 1.45 ± 0.13) compared with the control group (7.57 ± 0.05 and 7.59 ± 0.06) at 30- and 45-days post-surgery, respectively. The depth of the bone defects (mm) decreased significantly (p < 0.05) in the treated group (2.26 ± 0.12 and 0.008 ± 0.002) compared with the untreated group (4.05 ± 0.05 and 2.16 ± 0.07) at 30- and 45-days post-surgery, respectively. Throughout the period of study, there was a significant increase (p < 0.05) in the radiographic density of the bone defects (px) in the treated group (474 ± 17.88) compared with that in the control group (619.6 ± 6.85). SEM results revealed complete closure of the bone defects in the treated group. Thus, implantation of bone defects with the CS/PVA/GO/nano TiO₂ hydrogel represents a promising bone graft substitute for accelerating bone healing.

Abstract
Objective This study’s aim was: (1) introduce the digital drying oven as a reproducible, controllable, and accurate
heating device for burn model creation. (2) Define the heating temperature appropriate for developing consistent
second and third-degree burn injuries in rats.
Results Burns appeared deeper with more distinct borders in groups (B) and (C) than in group (A). The stainless-steel
rod at 100 ºC created burn injuries of the second degree, evidenced by the sloughing of the epidermis and necrosis
in the epithelium and upper part of the dermis. Heating at 150 and 200 ºC created third-degree burn injuries, where
necrosis involved the epidermis and dermis and extended to the subcutaneous fat and muscles. The depth of the
burn wound in the group (B) (371.2 ± 41.3 μm) and (C) (385.2 ± 38.0 μm) was significantly deeper compared with the
group (A) (178 ± 46.6 μm) (P < 0.001). The digital drying oven is a reliable, reproducible, and controllable heating device
for creating burn models. The stainless-steel rod (63 g and 8 mm) heated at 100 and 150 ºC with a contact time of 30 s
is adequate for creating consistent second and third-degree burn injuries in rats, respectively.

Egg production, one of the most important economic traits of chickens, is largely regulated by interactions of
genetic, endocrine, and environmental factors. Gut microbiota is the main environmental factor that is closely
related to egg production. However, the key gut microbiota affecting egg production is still unknown. Therefore,
Golden Montazah (GM) chickens (Rhode Island Red × Dokki-4 chickens) of 44 weeks of age were housed
separately. After keeping track of egg laying for 90 consecutive days, 10 laying hens were selected for the H
group (with a higher egg laying rate) based on the laying level, and 10 laying hens were assigned to the L group
(with a lower egg laying rate). Grossly, the number of eggs (P < 0.0001) and the number of hierarchical follicles
(P < 0.001) were significantly higher in the ovaries of high egg production chickens. HE staining results indicated
that the granulosa cell thickness of large white follicles (LWFs) was significantly higher (P < 0.001) in high
egg production chickens. 16S rRNA gene sequencing results showed that the relative abundance of Firmicutes
was higher both in the ileum and cecum of high egg production chickens, yet the relative abundance of Proteobacteria was higher in the ileum of low egg production chickens. Further, Spearman correlation analysis
indicated that the relative abundance of cecal Lachnoclostridium was positively correlated with the egg number
and hierarchical follicle number, while the relative abundance of ileal Olsenella was significantly (P < 0.05) and
positively correlated with the egg number. Yet the relative abundance of cecal Collinsella was significantly (P <
0.05) and negatively correlated with the egg number. Our findings indicated that gut microbiota is associated
with the egg-laying performance of chickens.

Pregnancy and lactation is a critical period for rabbit production. Estrogen (E2) and estrogen receptors
alpha (ERA) are essential during pregnancy and lactation and their importance stems from their
role in ovarian activities. Despite extensive research into the roles of E2 and its receptors in the
ovary, cellular distribution of ERA in the rabbit ovary during pregnancy, after parturition and during
lactation remained unexpectedly elusive. To achieve this aim, eighteen healthy sexually mature New
Zealand white rabbit does (2.97 ± 0.2 kg) were raised in the animal house, faculty of medicine, Assiut
University. The females rabbit were mated by fertile bucks; the day of mating as was considered Day 0
of pregnancy. Ovaries were collected at 12 h, 3, 7, 14 days post-mating, at parturition and at 10 days of
lactation and fixed then processed for immunohistochemistry of ERA. In the present study, the cellular
distribution of ERA in the rabbit ovary during pregnancy, postpartum and during lactation revealed
moderate ERA immunolocalization in the ovarian surface epithelial cells, stroma cells, fibroblast cells
of the tunica albuginea, and follicular cells of the primordial and primary follicles. The growing and
small antral follicles showed strong cytoplasmic and nuclear ERA immunolocalization in the granulosa
cells and theca folliculi cells. The large antral (graafian) and pre-ovulatory follicles showed moderate to
strong ERA immunolocalization in the granulosa cells, corona radiata cells, cumulus oopherous cells,
oocyte, theca interna cells and theca externa cells. The atretic antral follicle showed strong cytoplasmic
and negative nuclear ERA immunolocalization in the apoptotic granulosa cells and strong cytoplasmic
and nuclear ERA immunolocalization in the proliferated theca interna cells. The endothelial cells of the
ovarian blood vessels, the interstitial gland cells and telocytes showed strong cytoplasmic and nuclear
ERA immunolocalization. The corpus luteum (CL) during pregnancy till parturition showed moderate to
strong ERA immunolocalization in the large lutein cells, small lutein cells and luteal endothelial cells.
The regressed CL in the rabbit ovary 10 days of lactation showed weak ERA immunolocalization in the
regressed large lutein cells and moderate cytoplasmic and negative nuclear ERA immunolocalization
in the small lutein cells. Interestingly, the rabbit ovary during lactation showed abundant interstitial
gland with strong ERA immunolocalization in the interstitial gland cells. This work highlights the
role of ERA in the ovulation, folliculogenesis, lutenization and luteal regression in the rabbit during
pregnancy and lactation which contribute to enhancing this animal’s reproductive success.