Detection of human Sarcocystosis using dried blood on filter papers: An Immunofluorescent Antibody Test.

Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats' muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis.

Detection of human Sarcocystosis using dried blood on filter papers: An Immunofluorescent Antibody Test

@#Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats’ muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis.

Protective effect of magnesium acetyltaurate against endothelin-induced retinal and optic nerve injury.

Vascular dysregulation has long been recognized as an important pathophysiological factor underlying the development of glaucomatous neuropathy. Endothelin-1 (ET1) has been shown to be a key player due to its potent vasoconstrictive properties that result in retinal ischemia and oxidative stress leading to retinal ganglion cell (RGC) apoptosis and optic nerve (ON) damage. In this study we investigated the protective effects of magnesium acetyltaurate (MgAT) against retinal cell apoptosis and ON damage. MgAT was administered intravitreally prior to, along with or after administration of ET1. Seven days post-injection, animals were euthanized and retinae were subjected to morphometric analysis, TUNEL and caspase-3 staining. ON sections were stained with toluidine blue and were graded for neurodegenerative effects. Oxidative stress was also estimated in isolated retinae. Pre-treatment with MgAT significantly lowered ET1-induced retinal cell apoptosis as measured by retinal morphometry and TUNEL staining. This group of animals also showed significantly lesser caspase-3 activation and significantly reduced retinal oxidative stress compared to the animals that received intravitreal injection of only ET1. Additionally, the axonal degeneration in ON was markedly reduced in MgAT pretreated animals. The animals that received MgAT co- or post-treatment with ET1 also showed improvement in all parameters; however, the effects were not as significant as observed in MgAT pretreated animals. The current study showed that the intravitreal pre-treatment with MgAT reduces caspase-3 activation and prevents retinal cell apoptosis and axon loss in ON induced by ET1. This protective effect of ET1 was associated with reduced retinal oxidative stress.

Loss of human leucocyte antigen class I and gain of class II expression are early events in carcinogenesis: clues from a study of Barrett's oesophagus.

BACKGROUND: Human leucocyte antigen (HLA) expression is altered in oesophageal carcinomas compared with normal tissue. It is unclear, however, whether this phenotype precedes malignant transformation or results as a consequence of it. AIM: To investigate HLA class I and II expression in Barrett's oesophagus and normal squamous oesophageal tissue. METHODS: Asian patients with Barrett's oesophagus (n = 64) and a control group (n = 60) with a normal oesophagus but without reflux symptoms were recruited using endoscopic and histopathological criteria. Tissue samples were stained with monoclonal antibodies specific for HLA-ABC, HLA-DR alpha chain or HLA-DP/DQ/DR, and scored semiquantitatively. The results of immunohistochemical staining were correlated with clinical and histopathological characteristics of patients. RESULTS: Marked expression of HLA-ABC was observed in 50% of Barrett's oesophagus sections as compared with 68.3% of controls (p = 0.038). HLA-DR staining was seen in 51.6% of Barrett's oesophagus samples versus 11.7% of controls (p<0.001). Expression of HLA-DP/DQ/DR was evident in 73.4% of oesophageal intestinal metaplasia tissue as opposed to 18.3% of controls (p<0.001). Importantly, a total loss of HLA-ABC and a concomitant gain of HLA-DP/DQ/DR expression were seen in 37.5% of patients with Barrett's oesophagus but in none of the controls (p<0.001). Interestingly, this phenotype was associated positively with dysplasia (adjusted p, p* = 0.031) but negatively with non-steroidal anti-inflammatory drug use (p* = 0.004). CONCLUSIONS: HLA class I expression is down regulated and class II expression is up regulated in Barrett's oesophagus. As these changes predate malignant transformation, altered major histocompatibility complex expression may be a key event in disease progression, possibly in facilitating evasion from immune surveillance.

Detection of amplified int-2/FGF-3 gene in primary breast carcinomas using differential polymerase chain reaction.

Amplification of int-2/FGF-3 gene was investigated by differential polymerase chain reaction (dPCR) in 440 archival primary breast carcinoma tissues. Of these, 23 were comedo ductal carcinoma in situ (DCIS), 18 were non-comedo DCIS, 41 were comedo DCIS with adjacent invasive ductal carcinomas, 19 were non-comedo DCIS with adjacent invasive ductal carcinomas, 270 were invasive ductal carcinomas, 33 were invasive lobular carcinomas, 21 were colloid carcinomas and 15 were medullary carcinomas. Int-2 was amplified in 22% (96/440) of the primary breast carcinomas. It was shown that int-2 was amplified in 13% (3/23) of the comedo DCIS, 17% (7/41) of the comedo DCIS and 29% (12/41) of the adjacent invasive ductal carcinomas, 26% (71/270) of the invasive ductal carcinomas, 18% (6/33) of the invasive lobular carcinomas, 10% (2/21) of the colloid carcinomas and 13% (2/15) of the medullary carcinomas. In contrast, int-2 was not amplified in non-comedo DCIS and invasive ductal carcinomas with adjacent non-comedo DCIS lesions. A significant association was observed between int-2 amplification in the in situ components and adjacent invasive lesion (P<0.05). All tumors with int-2 amplification in the in situ lesions (7/7) also demonstrated same degree of amplification in the adjacent invasive components. However, 9% (5/53) of the tumors with no amplified int-2 gene in the in situ components showed int-2 amplification in the adjacent invasive lesions. A significant relationship was noted between amplification of int-2 and lymph node metastases (P<0.05) and poorly differentiated tumors (P<0.05) but not with estrogen receptor status (P>0.05) and proliferation index (Ki-67 and PCNA) (P>0.05). In Malaysia, majority of the patients belong to younger age group (<50 years old) but a comparison of the age groups showed that the amplification of int-2 was not statistically associated with patient age (P>0.05). These observations indicate that amplification of int-2 tends to strengthen the view that int-2 may have the potential to be an indicator of poor prognosis regardless of the age of the patient. Moreover, the presence of int-2 amplification in preinvasive, preinvasive and adjacent invasive lesions, and invasive carcinomas suggest that int-2 could be a marker of genetic instability occurring in early and late stages of tumor development.

Expression of c-erbB3 protein in primary breast carcinomas.

Expression of c-erbB3 protein was investigated in 104 primary breast carcinomas comprising nine comedo ductal carcinoma in situ (DCIS), 91 invasive ductal carcinomas and four invasive lobular carcinomas using two monoclonal antibodies, RTJ1 and RTJ2. Of the 91 invasive ductal carcinomas, seven contained the comedo DCIS component adjacent to the invasive component. An immunohistochemical technique was used to evaluate the association between expression of c-erbB3 and clinical parameters and tumour markers such as epidermal growth factor receptor (EGFR), c-erbB2, cathepsin-D and p53 in archival formalin-fixed paraffin-embedded tumour tissues. Our results indicated that RTJ1 and RTJ2 gave identical staining patterns and concordant results. It was found that the overexpression of c-erbB3 protein was observed in 67% (6/9) of comedo DCIS, 52% (44/84) of invasive ductal carcinomas, 71% (5/7) of carcinomas containing both the in situ and invasive lesions and 25% (1/4) of invasive lobular carcinomas. A significant relationship (P < 0.05) was observed between strong immunoreactivity of c-erbB3 protein and histological grade, EGFR and cathepsin-D, but not with expression of c-erbB2, p53, oestrogen receptor status, lymph node metastases or age of patient. However, we noted that a high percentage of oestrogen receptor-negative tumours (59%), lymph node-positive tumours (63%) and c-erbB2 (63%) were strongly positive for c-erbB3 protein. We have also documented that a high percentage of EGFR (67%), c-erbB2 (67%), p53 (75%) and cathepsin-D-positive DCIS (60%) were strongly positive for c-erbB3. These observations suggest that overexpression of c-erbB3 protein could play an important role in tumour progression from non-invasive to invasive and, also, that it may have the potential to be used as a marker for poor prognosis of breast cancer.

Unusual presentation of cancer.

The large bowel is the most frequent primary site for metastases in inguinal hernial sacs. We report four cases, two due to carcinoid of unknown primary, and one each due to adenocarcinoma of colon, stomach and pancreas. We recommend that all hernial sacs, particularly in the elderly, be examined microscopically.

Differentiating medulloblastoma in adults.

We present two cases of medulloblastoma in adults patients showing extensive astrocytic differentiation. In one case, there was synchronous oligodendroglial differentiation, both changes appearing after treatment with radiotherapy. The possible role of radiotherapy in the differentiation is discussed.

The skeletal manifestations of brucellosis.

A prospective study of 33 patients with skeletal complications of brucellosis is reported. The diagnosis depends on the brucellar agglutination titre because the clinical, radiological and histological features are not specific, and blood culture and culture of synovial fluid are often negative. The commonest site is the sacroiliac joint, and brucellosis must be differentiated from tuberculosis as an important cause of backache. Treatment is with chemotherapy, the preferred combination being rifampicin and co-trimoxazole. Relapse can occur, but the chances can be reduced by careful clinical monitoring, and by repeated serological tests and gallium scanning; it is treated by further chemotherapy. Operation is indicated to relieve spinal cord compression or to drain a large paravertebral abscess.

Nasopharyngeal carcinoma. A comparative study.

A comparative study of Ghanaian and Saudi hospital cases of nasopharyngeal carcinoma (NPC) shows a higher occurrence among younger subjects in the Ghanaian group, with the peak age incidence in the Saudi group being similar to those reported from other parts of the world. In view of the known high prevalence of Burkitt's lymphoma among Ghanaian children, it is suggested that the much higher proportion of younger Ghanaian subjects with NPC may be related to early exposure to the Epstein-Barr virus which is known to be associated with both malignancies.

Histiocytic necrotising lymphadenitis (Kikuchi-Fujimoto disease) in Saudi Arabia.

Five cases of histiocytic necrotising lymphadenitis (Kikuchi-Fujimoto disease) are described. As the condition appears to be poorly recognised in Saudi Arabia, an extensive review of the histological features is presented, in order to improve pathologists' awareness and to minimise the risk of diagnosing prognostically less favourable conditions such as malignant lymphoma. Etiologic and pathogenetic features of the disease are also discussed.

Dissemination in cutaneous leishmaniasis. 3. Lymph node involvement.

In a study of zoonotic cutaneous leishmaniasis (CL) due to leishmania major in an endemic focus in Saudi Arabia, lymph node enlargement was observed in 66 of 643 patients (10.26%). The epitrochlear lymph nodes were most commonly involved (68%), but cervical (11%), axillary (15%), and inguinal (18%) lymph nodes were also involved. In eight patients (12%), two lymph node areas were involved. The affected lymph nodes were typically solitary, firm, mobile, nontender, only moderately enlarged, and appeared to persist beyond the clinical healing of the associated skin lesions. The results of pathologic and immunopathologic studies carried out on eight lymph nodes obtained from this group of patients supported the leishmanial etiology. Although amastigotes were only demonstrated in two lymph nodes, the leishmanial antigen was found in all eight specimens. The lymph node involvement is another manifestation of dissemination in infection with a dermotropic leishmania. It appears that in some cases, instead of the parasite, it may be the leishmanial antigen that disseminates and produces lymphadenitis.

Dissemination in cutaneous leishmaniasis. II. Satellite papules and subcutaneous induration.

In a field study of cutaneous leishmaniasis (CL) due to Leishmania major, zymodeme Lon-4, in an endemic focus in Saudi Arabia, 80 patients in a group of 643 patients (12.44%) were found to have multiple, inflammatory, satellite papules (SP) around one or more CL lesions. The SP often appeared to erupt after commencement of antileishmanial treatment. They showed a range of morphology and pathology, and comparisons with corresponding CL lesions showed important differences. Amastigotes were seen in only one of seven biopsy specimens. Another morphologic feature, subcutaneous induration (SCI), was noted on routine palpation in 20 patients in the same patient group (3.11%). The SCI either was radiating all around the lesion and appeared as an "iceberg nodule" or was present as a "tonguelike" process proximal to the CL lesion. Amastigotes were seen in four of nine biopsy specimens of SCI. SP and SCI represent reactions to local dissemination of the parasite or its antigenic products, and as morphologic features aid in the clinical diagnosis of CL.

Progressive subluxation of the wrist in sickle-cell disease.

Progressive subluxation of the wrist as a sequel to vasoocclusive crisis of sickle-cell disease is described. This shares many features with Madelung's deformity. The management depends on the functional disability and the cosmetic appearance. One of our patients needed surgical correction to improve the deformity and the range of motion.

Septic arthritis in sickle cell disease.

We report a prospective study of fourteen patients with sickle cell disease who were suffering from septic arthritis. They made up 11% of 129 patients with orthopaedic complications of the disease. The infecting organism in eleven was a staphylococcus, with salmonella in only two. The severity of the condition and its sequelae are described. The best treatment is open surgical drainage and intensive chemotherapy.

Peripheral nerve involvement in cutaneous leishmaniasis (Old World).

A review of 288 skin biopsy specimens from cutaneous leishmaniasis lesions caused by Leishmania major showed assorted nerve changes in 14 biopsy specimens (5%). Ten patients had perineural inflammatory cell infiltrate consisting of either lymphocytes or a mixture of lymphocytes, plasma cells, and macrophages. Four patients had inflammatory cell invasion of the nerves (neuritis), and in one of them the inflammation was granulomatous and associated with nerve destruction. Amastigotes were seen inside the nerves in two patients. Sensory testing of 50 consecutive patients with cutaneous leishmaniasis identified two patients with diminished sensations over the lesions.

Dissemination in cutaneous leishmaniasis. I. Subcutaneous nodules.

In a study of cutaneous leishmaniasis (CL) caused by Leishmania major in Saudi Arabia, 10% of the patients were found to have subcutaneous nodules (SCN). The SCNs were usually inconspicuous, painless, and proximal to the primary skin lesions; when multiple, they showed a "sporotrichoid" configuration or appeared as "beaded cords." Their number ranged from 1-16 (average 3 25 +/- 2.50; mean +/- 1 SD). In some patients, the SCNs seemed to be triggered by antileishmanial treatment. The clinical picture and pathologic findings suggest that SCNs in patients with CL represent lymphatic dissemination, a phenomenon not widely recognized.

Immunohistological investigations in chronic cutaneous leishmaniasis in Saudi Arabia.

The distribution and numbers of IgG-, IgA-, IgM-, and lysozyme-positive cells were investigated by the immunoperoxidase method in paraffin-sections of 13 cases of chronic cutaneous leishmaniasis of low parasite load from Saudi Arabia. The majority of the peroxidase-positive plasma cells contained IgG, whereas the numbers of IgA+ and IgM+ plasma cells were not so numerous. Small groups of squamous epithelial cells showed immunoreactivity for IgG and IgA. Similar positive staining was observed extracellularly in the oedematous upper dermis, in the endothelial cells, and in the perivascular space. The activated macrophages showed strong and diffuse peroxidase staining for lysozyme, whereas epithelioid cells and multinucleated giant cells were negative or had finely granular and considerably weaker staining. It is suggested that humoral immunity also participates in the elimination of the parasites and an immunologically induced necrosis might be responsible for the ulceration of the skin in cutaneous leishmaniasis. It is also assumed that the lysozyme immunoreactivity can be a marker of the activation state of the macrophages.

Alimentary malignancies in Arabs in the Eastern Province of Saudi Arabia.

A study was made of 89 cases of alimentary malignancies in Arab patients in the Eastern Province of Saudi Arabia. The pattern of alimentary involvement observed was colo-rectum 43%, stomach 34%, oesophagus 19% and small intestine 4.5%. This finding was in contrast to previous studies in Saudi Arabia which had reported the upper alimentary tract, particularly the oesophagus, as the site most frequently affected. Of the patients 80% were Saudis. The male:female ratio was 3.9:1. A striking feature was the youthfulness of several patients, 44% being under fifty years of age. There were only three cases of alimentary lymphoma. Although colo-rectal schistosomiasis was a moderately frequent finding in the population studied, there was no pathological evidence to support an aetiological association between bilharziasis and large bowel malignancy.