Presence of Lipid Vacuoles in Kupffer Cells as a Marker for Early Detection of Niemann-Pick Disease Type C in Neonatal Cholestasis

Background Early diagnosis of Niemann-Pick disease type C (NP-C) is the key for reduction of organ damage since a medical treatment is available now. However, early detection of patients with a high clinical suspicion of NP-C in neonatal cholestasis is still challenging. Plasma oxysterol studies demonstrate a relatively low specificity for NP-C in neonatal cholestatic patients. This study explores the significance of lipid vacuoles in Kupffer cells for early detection of NP-C in neonatal cholestasis. Results Between January 2018 and December 2020, 168 neonatal cholestatic patients with unexplained causes were underwent both liver biopsy and genetic tests, and 26 detected lipid vacuoles in Kupffer cells by CD68 staining. NP-C was diagnosed in 6 out of the 26 patients (23.1%), comparing to none of the remaining 142 neonatal cholestatic patients without vacuolar Kupffer cells (P < 0.001). In neonatal cholestatic patients with vacuolar Kupffer cells plus splenomegaly, the ratio of positive diagnosis of NP-C was 31.6% (6/19). Between January 2015 and December 2017, 3 additional NP-C patients were diagnosed. Of them, 1 patient underwent liver histologic studies also revealing lipid vacuoles in Kupffer cells. The 7 NP-C patients underwent liver biopsy at ages ranging from 35 to 112 days. Lipid vacuoles were only detected in a few Kupffer cells in the early disease course (about 1 month of age). The amount of vacuolar Kupffer cells increased with age, and the size became enlarged. Typical foam cells were only observed in HE sections of liver tissues obtained beyond 3 months of age when enlarged Kupffer cells with lipid vacuoles became obvious. In addition, 4 out of the 9 NP-C patients had fasting hypoglycemia. Conclusions Lipid vacuoles in Kupffer cells from liver biopsy can serve as a screening marker for early detection of NP-C in neonatal cholestasis.

Sporadic hemangioblastoma of the kidney: a clinicopathologic study of three cases and a literature review

Much attention has been paid to renal hemangioblastoma, but there are still challenges in its differential diagnosis. Three cases (2 men, 1 woman; age: 40–56 years) presented with renal tumors. The tumors were surrounded by a thick fibrous capsule, well-demarcated from the surrounding renal parenchyma, and composed of sheets or nests of polygonal to short spindle-shaped tumor cells with a rich capillary network. In cases 1 and 3, the large polygonal tumor cells contained abundant pale or eosinophilic cytoplasm, and some possessed intracytoplasmic lipid vacuoles. In case 2, tumor cells were characterized by a uniform size, mild, clear, or lightly stained cytoplasm, and typical "clear cell" appearance. Immunohistochemistry revealed that the polygonal stromal cells were strongly and diffusely positive for α-inhibin, neuron-specific enolase (NSE), S100 protein, and vimentin. Cluster of differentiation (CD)10 and paired box gene (PAX)8 were positive, while epithelial membrane antigen (EMA) and cytokeratin (CK) were focally positive in case 3. CD34 and CD31 outlined the contours and distribution of the vascular networks. Renal hemangioblastoma is rare and prone to misdiagnosis; more attention should be paid to the morphological features and reasonable application of immunohistochemistry in the diagnosis of hemangioblastoma.

Primary Splenic Pleomorphic Liposarcoma in a Bitch

Background: Liposarcoma is a malignant neoplasm of lipoblasts with low incidence in dogs, representing 1.7% of neoplasms diagnosed in the spleen. In veterinary medicine, this neoplasm is classified morphologically into the myxoid, well-differentiated, undifferentiated and pleomorphic subtypes, the latter being one of the most aggressive forms, mainly in cavity organs. This study reports a case of primary splenic pleomorphic liposarcoma in a female dog, addressing anatomopathological and immunohistochemical aspects.Case: A 14-year-old, 35 kg female mongrel canine with a history of absence of defecation, progressive weight loss, difficulty walking, sensitivity to abdominal palpation, prostration, pale mucous membranes, tachypnea and abdominal distention. The condition evolved to death and, on necroscopy, there was an increase in splenic volume with neoformation of whitish and reddish color, measuring 32 × 27 cm in its largest axes and weighing 8.9 kg. The neoformation exhibited areas of firm and soft consistency, and sectioning revealed the existence of focal areas of extensive necrosis and cavity collections of different diameters that allowed the flow of liquid serous contents with a brownish red color. Microscopy showed cells of neoplastic morphology infiltrating the splenic parenchyma, mostly with slightly acidophilic cytoplasm and few intracytoplasmic lipid vacuoles, which varied in size and distribution. The nuclei of the cells were large, eccentric and irregular, with round to oval morphology, grossly lacy chromatin and single or multiple evident nucleoli. These cells exhibited marked anisocytosis, anisokaryosis and pleomorphism, with more than one mitotic figure per high magnification field visible. Moderately inflammatory infiltrate, predominantly lymphocytic, permeated the neoplastic cells, and marked depletion of lymphoid follicles and atrophy of the red pulp were found in the remaining splenic parenchyma. Immunohistochemical tests revealed marked and discrete immunostaining for anti-vimentin and anti-S100 antibodies, respectively. No staining was observed for anti-pan cytokeratin, anti-desmin, anti-alpha smooth muscle actin or anti-CD20 antibodies. Based on anatomopathological and immunohistochemical aspects, it was concluded to be a splenic pleomorphic liposarcoma of primary origin.Discussion: the spleen is not a common anatomical site for the development of liposarcoma, a neoplasm whose origin remains unclear. Similar to what occurs in humans, liposarcoma is believed to develop from the adipose tissue of the splenic hilum. Thus, it should be considered as a differential diagnosis of invasive abdominal tumors. For the identification and classification of liposarcoma as a pleomorphic subtype, we considered mainly histological findings such as marked cell atypia and intracytoplasmic lipid vacuoles, which may or may not be present in neoplastic cells. Immunohistochemical examination favored the diagnosis of liposarcoma, regardless of the subtype, due to the marked immunostaining for the anti-vimentin antibody, unlike immunostaining for the anti-S100 antibody, for which it was variable. This fact is related to adipocyte differentiation, where lower amounts of intracytoplasmic lipids translate into lower immunostaining intensity for anti-S100. Histological and immunostaining aspects should be regarded with caution in the diagnosis of pleomorphic liposarcoma, as it is a distinct neoplastic entity, with a complex karyotype and without correlation with the other subtypes.

Sporadic Hemangioblastoma of the Kidney: A clinicopathologic study of 3 cases and review of the literature

Background: Hemangioblastoma is a benign tumor of unknown histogenesis that mainly occurs in the central nervous system (CNS) associated with von Hippel-Lindau (VHL) disease. Much attention has been paid to the renal hemangioblastoma, but there are still some challenges in the differential diagnosis.Case presentation: Here, we describe three cases of sporadic renal hemangioblastoma with no clinical features of VHL diseases. All the three patients (male: 2; female: 1) were 40-56 years old. In all cases, the tumors were surrounded by a thick fibrous capsule and well-demarcated from the surrounding renal parenchyma. Tumors were composed of sheets or nests of polygonal to short spindle tumor cells and a rich capillary network. In case 1 and case 3, the large polygonal tumor cells contained abundant pale or eosinophilic cytoplasm, and some of the cells possessed intracytoplasmic lipid vacuoles. In case 2, tumor cells were characterized by uniform size, mild, clear or lightly stained cytoplasm and typical "clear cell" appearance. In the views of immunohistochemistry, the polygonal stromal cells were strongly and diffusely positive for α-inhibin, NSE, S100 protein, and vimentin. CD10 and PAX8 were positive, while EMA and CK showed focally positive in case 3. CK8/18, HMB45, MelanA, CgA, Syn, SMA, Desmin and CD56 were all negative. CD34 and CD31 outlined the contours and distribution of vascular networks in tumors. Conclusions: Renal hemangioblastoma is rare and prone to be misdiagnosed. More attention should be paid to the morphological features and reasonable application of immunohistochemistry for the diagnosis of hemangioblastoma.

The Expression Profile of Dental Pulp-Derived Stromal Cells Supports Their Limited Capacity to Differentiate into Adipogenic Cells

Mesenchymal stromal cells (MSCs) can self-renew, differentiate into specialised cells and have different embryonic origins—ectodermal for dental pulp-derived MSCs (DPSCs) and mesodermal for adipose tissue-derived MSCs (ADSCs). Data on DPSCs adipogenic differentiation potential and timing vary, and the lack of molecular and genetic information prompted us to gain a better understanding of DPSCs adipogenic differentiation potential and gene expression profile. While DPSCs differentiated readily along osteogenic and chondrogenic pathways, after 21 days in two different types of adipogenic induction media, DPSCs cultures did not contain lipid vacuoles and had low expression levels of the adipogenic genes proliferator-activated receptor gamma (PPARG), lipoprotein lipase (LPL) and CCAAT/enhancer-binding protein alpha (CEBPA). To better understand this limitation in adipogenesis, transcriptome analysis in undifferentiated DPSCs was carried out, with the ADSC transcriptome used as a positive control. In total, 14,871 transcripts were common to DPSCs and ADSCs, some were unique (DPSCs: 471, ADSCs: 1032), and 510 were differentially expressed genes. Detailed analyses of overrepresented transcripts showed that DPSCs express genes that inhibit adipogenic differentiation, revealing the possible mechanism for their limited adipogenesis.

Effect of dietary resveratrol on cell-mediated immunity and hepatocyte morphometry in the model organism medaka (Oryzias latipes Temminck & Schlegel)

The effect of dietary resveratrol (40 µg g−1 BW day−1) on cell-mediated immunity (the activity of spleen phagocytes and the proliferative response of lymphocytes) and liver histology (hepatocyte morphometry, lipid vacuoles, and glycogen granules) in adult medaka (Oryzias latipes) (aged two years, body weight BW = 0.49 g) were tested after ten days of feeding with experimental diets. The fish were fed either a commercial diet (control group C) or this same diet supplemented with resveratrol (group R) three times daily at 3% of body weight (BW). Fish fed feed with resveratrol had significantly higher intracellular phagocyte killing activity than did those in the control group (P < 0.05). No differences in the sizes of hepatocytes or their nuclei were observed in the fish from groups C and R. The nucleo-cytoplasmic ratio ranged from 0.39 (± 0.03; group R) to 0.42 (± 0.03, group C) in adult medaka exposed to the two dietary treatments (P > 0.05). The results indicated that the diet supplemented with resveratrol at a dose of 40 µg g−1 BW day−1 had no impact on the liver tissues. Typical hepatocytes with lipids and glycogen were observed to the same extent in the vacuoles. Moreover, no mortality or pathological changes were noted in the fish studied. The diet containing resveratrol helped to maintain disease resistance without affecting the liver tissue after ten days of feeding.

Cystic neutrophilic granulomatous mastitis: an update

Cystic neutrophilic granulomatous mastitis (CNGM) is a rare subtype of granulomatous mastitis with a highly distinct histological pattern often associated with Corynebacterium species. CNGM is characterised by suppurative lipogranulomas that are composed of central lipid vacuoles rimmed by neutrophils and an outer cuff of epithelioid histiocytes. Some of the lipid vacuoles may contain sparse, rod-shaped, gram-positive bacilli that can be easily missed or dismissed. The surrounding mixed inflammatory infiltrate contains Langhans-type giant cells, lymphocytes and neutrophils. CNGM occurs in reproductive age women with a history of pregnancy and typically presents as a palpable mass that can be painful. CNGM has many mimickers, most significantly breast carcinoma. In many cases, CNGM has significant pathological and clinical overlap with other forms of granulomatous mastitis. Given the association with Corynebacterium species, early diagnosis of CNGM is essential in offering patients the most appropriate treatment. Prolonged antibiotic therapy specifically directed to corynebacteria is required, sometimes even beyond resolution of clinical symptoms. This comprehensive review of the existing literature on CNGM describes clinical–pathological features, microbiological findings, challenges associated with the microscopic differential diagnosis, clinical implications of this diagnosis and emerging treatment options. Morphological criteria and suggested comments to convey the degree of diagnostic certainty are also proposed for standard pathology reporting.

Structural Organization of the Liver in the Presence of Pig Hepatosis

The structural organization of the liver in hepatosis and its clinical and morphological manifestation in pigs were studied. In animals with reduced vitality, the liver underwent deep microscopic and ultrastructural changes. At hepatic steatosis, the liver had a cellular structure, many lipid vacuoles and the damage of mitochondrial membranes of a granular endoplasmic reticulum appeared in hepatocytes. Proteinosis was characterized by impaired of lobular structure, with protein swelling of the cytoplasm of hepatocytes and Disse’s spaces expansion, significant decrease of glycogen; mitochondrial swelling and the appearance of lysosomal vacuoles in the hepatocyte. In massive hepatic necrosis, hepatocyte dystrophy was noted, significant histochemical decrease of glycogen, lysis of the hepatocyte granule endoplasmic reticulum, pycnosis of the hepatocyte nucleus, and in general necrobiotic changes in the liver cells.

Effect of Tff3 Deficiency and ER Stress in the Liver

Endoplasmic reticulum (ER) stress, a cellular condition caused by the accumulation of unfolded proteins inside the ER, has been recognized as a major pathological mechanism in a variety of conditions, including cancer, metabolic and neurodegenerative diseases. Trefoil factor family (TFFs) peptides are present in different epithelial organs, blood supply, neural tissues, as well as in the liver, and their deficiency has been linked to the ER function. Complete ablation of Tff3 expression is observed in steatosis, and as the most prominent change in the early phase of diabetes in multigenic mouse models of diabesity. To elucidate the role of Tff3 deficiency on different pathologically relevant pathways, we have developed a new congenic mouse model Tff3−/−/C57BL6/N from a mixed background strain (C57BL6/N /SV129) by using a speed congenics approach. Acute ER stress was evoked by tunicamycin treatment, and mice were sacrificed after 24 h. Afterwards the effect of Tff3 deficiency was evaluated with regard to the expression of relevant oxidative and ER stress genes, relevant proinflammatory cytokines/chemokines, and the global protein content. The most dramatic change was noticed at the level of inflammation-related genes, while markers for unfolded protein response were not significantly affected. Ultrastructural analysis confirmed that the size of lipid vacuoles was affected as well. Since the liver acts as an important metabolic and immunological organ, the influence of Tff3 deficiency and physiological function possibly reflects on the whole organism.

The Effect of Dayak Onion Bulb Ethanol Extract (Sisyrinchium palmifolium L.) on Triglyceride Level and Aorta Histopathology in Diabetes Melitus White Rat Induced by Alloxan

The high prevalence of diabetes mellitus in Indonesia has made antidiabetic treatment efforts increasingly popular. In addition, the many side effects caused by the use of chemical drugs, triggered the development of research on herbal therapy. The purpose of this study was to determine the effectiveness of Dayak onion bulb extract in reducing two-hour postprandial blood glucose, triglyceride levels, and the amount of lipid vacuoles in aorta blood vessels of diabetic rat. The long-term goal of this study is the creation of innovation products in the form of standardized herbs as complementary therapies that can be used daily by the community, so that the risk of morbidity can be reduced. The research design used was randomized control group pretest posttest design. The results of blood glucose analysis showed that there were significant differences between the positive control group with the 400 mg/KgBW and 800 mg/KgBW extract group, with a significance value of 0,000. In the extract group, 400 mg/KgBW with 800 mg/KgBW extract group showed no significant difference with a significance value of 0.390. The results of the analysis of triglyceride levels showed no significant difference between the positive control group with the 400 mg/KgBW extract group, with a significance value of 0.981. However, there were significant differences between the positive control group with the 800 mg/KgBW extract group with a significance value of 0.025. Between the 400 mg/KgBW and 800 mg/KgBW extract groups, there was a difference with a significance value of 0.024. Aorta histopathology results showed that there was a significant difference in the number of lipid vacuoles between the positive control group and the 400 mg/KgBW extract group and 800 mg/KgBW extract group. Based on the research, it can be concluded that the extract dose of 400 mg/KgBW is a dose that is able to influence the decrease in blood glucose levels, triglycerides and the amount of lipid vacuoles in the aorta blood vessels.