Growth characteristics and cytokine/chemokine induction profiles of dengue viruses in various cell lines.

The dengue disease varies clinically from mild fever to hemorrhagic fever up to potentially fatal shock syndrome. Such differences in manifestation of dengue virus (DENV) infection have been ascribed to intrinsic differences in DENVs. Four DENV serotypes or four dengue viruses (DENV-1 to DENV-4) have been clearly distinguished; however, only limited information is available on their biological characteristics in vitro. To shed more light on this subject, replication kinetics of all four DENVs in various cell lines and induction of 26 cytokines/chemokines were investigated. The results showed that, (i) all DENVs replicated relatively similarly in various cell lines, (ii) DENV-1 was most effective in this regard, (iii) A549 cells showed the highest virus replication rate compared to other cells, and (iv) all DENVs induced in A549 cells similar levels of several cytokines/chemokines, namely eotaxin, granulocyte colony stimulating factor (G-CSF), interferon alpha 2 (IFN-α2), interleukins 6, 8 15 (IL-6, IL-8, IL-15), and IP-10. In conclusion, this study revealed a similarity in growth characteristics and cytokine/chemokine induction profile for all four DENV serotypes in vitro.

The role of anti-dengue virus NS-1 and anti-protein disulfide isomerase antibodies on platelet aggregation in secondary dengue infection.

AIM: to observe the correlation between anti-NS-1 and anti PDI antibodies against platelets function disorder on secondary dengue infection. METHODS: 50 patients with secondary DV infection according to WHO criteria were observed by a cross sectional study. Patient's blood was collected on day 3, 5 and 7 after fever onset. Platelets aggregation test was done to prove the possibility of platelets dysfunction. Anti-NS-1 and anti-PDI antibodies were determined by solid phase ELISA. RESULTS: the inhibition of platelets aggregation was increased among day of observation. Means value of inhibition on day 3 is 46.6%, day 5 is 52.5% and day 7 is 56%. There is a significant difference (p<0.05) of inhibition of platelet aggregation value between days of observation. The antibodies against NS-1 DV and PDI were detected in all 50 sera with the positive rate of 90% develop NS-1 antibodies and 72% of PDI antibodies, on day 3 of symptoms. The highest OD of NS-1 antibodies is detected on the day 3 and decreased on day 7. The OD of PDI antibodies was increased on day 3 and still increasing on day 7. There is a significant correlation between anti NS-1 and PDI antibodies (r=0.386-0.490), while the differences of OD between observation days are not significant (p>0.05). CONCLUSION: the kinetics profile of NS1 and PDI antibodies responses, which were detected by the third day of symptoms. Dengue patients' sera inhibited platelets aggregation. NS-1 antibodies and PDI antibodies might have a role on the platelets aggregation dysfunction; however, there is no correlation between them. It is possible that other mechanism involve in the inhibition of platelets aggregation.

Hepatosplenic T-cell lymphoma presented with massive splenomegaly and pancytopenia - a case report.

BACKGROUND: Hepatosplenic T-cell lymphoma (HSTCL) is a rare subtype of peripheral T-cell lymphoma. Patients usually present with splenomegaly and pancytopenia but without lymphadenopathy. Immunohistochemistry (IHC) staining of bone marrow biopsy shows intra-sinusoidal infiltration of CD3 and CD56 T-lymphocytes. Current treatment strategy of HSTCL includes a CHOP regimen (cyclophosphamide, adriamycine, vincristine, prednisone) followed by autologous transplantation. CASE: A 28-year-old male presented with abdominal fullness, weight loss, and massive splenomegaly. Laboratory findings revealed pancytopenia. A CT scan of the abdomen displayed hepatomegaly and massive splenomegaly. The bone marrow pathology examination showed monotonous medium-sized lymphocytes with some cluster of atypical lymphocytes with loosely condensed chromatin and pale cytoplasm. The intra-sinusoidal location was more prominent after using IHC staining of CD3 and CD56, which are characteristics of HSTCL. We administered CHOP-based regiment every 3 weeks for 3 cycles; however, the response was a stable disease. Since the splenomegaly was still massive and compromised the patient, the multidisciplinary team decided to perform splenectomy. Unfortunately, the patient did not survive the surgery. CONCLUSION: Hepatosplenic T-cell lymphoma is a rare aggressive disease, which is part of peripheral T-cell lymphoma. CHOP-based chemotherapy appeared to be ineffective, and we need further studies to find the optimal treatment of HSTCL.

Sulawesi propolis induces higher apoptotic activity and lower inflammatory activity in a rat endometriosis model.

Background: Endometriosis has a major impact on women's quality of life. The two primary pathologies are chronic inflammation and altered apoptotic activity. Sulawesi propolis has been shown to have known anti-inflammatory and pro-apoptotic properties in other diseases. Objective: To investigate the effects of Sulawesi propolis in the rat endometriosis model. Methods: An autologous endometriosis model was created in 60 female Wistar rats by laparotomy. Rats were divided into four groups (n = 15 in each group): control group (CG), dienogest group (DG), propolis 50 mg/kg body weight (BW)/day (P50) group, and propolis 100 mg/kg BW/day (P100) group. Each treatment group was divided into three different treatment durations (n = 5 in each treatment group): 2, 4 and 6 weeks. After treatment, laparotomy was performed to determine endometriotic tissue growth, apoptosis [caspase-3 and Bcl-2-associated X/Bcl-2 (Bax/Bcl)] and inflammation [prostaglandin-E2 (PGE2) and interleukin-1B (IL-1B)]. Results: A significant difference was seen in endometriotic tissue growth between the P50 group and the CG, with the greatest reduction in the P50 6-week (P50-6) group, reaching 70.66% of the initial area. Highest Bax/Bcl-2 mRNA expression was shown in the P50-4 and P100-4 groups, highest caspase-3 expression was shown in the P50-2 and P50-4 groups, and lowest IL-1B expression was shown in the P50-4 group; all differed significantly from the CG. No significant difference in PGE2S mRNA was found between the groups. Conclusion: Sulawesi propolis extract suppressed endometriotic tissue growth in the rat model by increasing apoptotic activity. The effects were time-dependent, with 50 mg/kg BW as the optimal dose.

Diffuse large B-cell lymphoma associated ileocecal intussusception in adulthood.

BACKGROUND: Intussusception in adults is considered a rare condition, accounting for 5% of all cases of intussusceptions and approx. 1-5% of bowel obstruction. Almost half intussusceptions of the bowel are associated with malignant disease; thus, we should also treat the underlying malignancy. CASE DESCRIPTION: A 52-year-old male presented with colicky right lower abdominal pain for a 6-month period. He had a weight loss of 20kg within 6 months. Physical examination revealed a tender right lower abdominal mass. Colonoscopy showed a mass that filled the ileocecal. The digestive surgeon performed laparoscopic right hemicolectomy with end-to-end anastomosis. Histopathology examination showed diffuse proliferation of large tumor cells with centroblastic-like features prominently in submucosal area, with normal epithelial mucosa. The immunohistochemistry result concluded the final dia-gnosis of diffuse large B-cell lymphoma. RCHOP chemotherapy regimens were administered every 3 weeks for 6 cycles. The response was complete remission. DISCUSSION: Intussusception was preoperatively dia-gnosed by multi-slice spiral CT scans with the characteristic target or sausage sign, edematous bowel wall and mesentery in the lumen. After surgery, approximately 90% of adult intussusception cases have a demonstrable etiology. Malignant lymphoma, especially diffuse large B-cell lymphoma, of the ileocecal is one cause of the adult intussusception. CONCLUSION: Adult bowel intussusception is a rare clinical entity. Abdominal CT is considered as the most sensitive imaging modality in the dia-gnosis of intussusception. Diffuse large B-cell lymphoma is the most common cause of ileocecal intussusception.

Primary breast lymphoma - a case report.

BACKGROUND: Primary breast lymphoma is a rare disease and accounts for 0.4-0.5% of malignant breast neoplasms and 1.7-2.2% of extra-nodal lymphomas, with diffuse large B-cell lymphoma (DLBCL) as the most common histologic subtype. CASE: A 47-year-old female with beta thalassemia presented with a lump of the left breast, redness, pain, and swelling of her left breast. Physical examination showed tender, red, swollen left breast. Laboratory findings show mild anemia and normal level of lactate dehydrogenase 329 U/L (normal range: 240-480 U/L). PET scan showed hypermetabolic mass with irregular margins covering the whole left breast quadrants with the size of 11.25 x 5.17cm with left pectoralis major, left parasternal, and left axillary hypermetabolic nodules. Histopathology and immunohistochemistry staining showed a non-germinal center B-cell-like subtype of DLBCL CD20+. We administered the R-CHOP regimen (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednison) every 3 weeks for 6 cycles. The response was complete remission. The patient tolerated the chemotherapy well and achieved long term complete remission. CONCLUSION: Primary breast lymphoma is a rare disease with the most common subtype is diffuse large B-cell lymphoma. Systemic chemother-apy R-CHOP is the treatment option for primary breast diffuse large B-cell lymphoma.

The evolutionary origins of Southeast Asian Ovalocytosis.

Southeast Asian Ovalocytosis (SAO) is a common red blood cell disorder that is maintained as a balanced polymorphism in human populations. In individuals heterozygous for the SAO-causing mutation there are minimal detrimental effects and well-documented protection from severe malaria caused by Plasmodium vivax and Plasmodium falciparum; however, the SAO-causing mutation is fully lethal in utero when homozygous. The present-day high frequency of SAO in Island Southeast Asia indicates the trait is maintained by strong heterozygote advantage. Our study elucidates the evolutionary origin of SAO by characterizing DNA sequence variation in a 9.5 kilobase region surrounding the causal mutation in the SLC4A1 gene. We find substantial haplotype diversity among SAO chromosomes and estimate the age of the trait to be approximately 10,005 years (95% CI: 4930-23,200 years). This date is far older than any other human malaria-resistance trait examined previously in Southeast Asia, and considerably pre-dates the widespread adoption of agriculture associated with the spread of speakers of Austronesian languages some 4000 years ago. Using a genealogy-based method we find no evidence of historical positive selection acting on SAO (s=0.0, 95% CI: 0.0-0.03), in sharp contrast to the strong present-day selection coefficient (e.g., 0.09) estimated from the frequency of this recessively lethal trait. This discrepancy may be due to a recent increase in malaria-driven selection pressure following the spread of agriculture, with SAO targeted as a standing variant by positive selection in malarial populations.

Molecular Basis of beta-Thalassemia in Indonesia: Application to Prenatal Diagnosis.

Background: To facilitate an effective prevention program, the beta-thalassemia mutations in the different ethnic groups in Indonesia were characterized. Methods and Results: The amplification refractory mutation system and artificially created restriction site were used to detect seven known mutations previously described in the Indonesian population. Other mutant alleles were identified by chemical cleavage mismatch, double-stranded sequencing, and Southern blotting. With these methods 78% of beta-thalassemia mutant alleles have been detected so far. Thirteen different beta-thalassemia mutations were characterized, nine of which had previously been described in the Jakarta population. The most frequent mutation is HbE (29%), followed by IVS1-nt5 (19%), and Cd 35 (8%). The frequencies of the other mutations varied from 4% to less than 1%. Two large gene deletions, Filipino beta-deletion and Hb Lepore, were identified in patients from the eastern part of Indonesia. Conclusions: The ethnicity and clinical hematology of cases in the region should be considered in the screening strategy for carriers and antenatal diagnosis of beta-thalassemia in Indonesia. Direct sequencing proved to be the appropriate method for detecting the unknown mutations, and Southern blotting had to be used for large deletions.

Detection of beta-globin gene mutations among Kelantan Malay thalassaemia patients by polymerase chain reaction restriction fragment length polymorphism.

INTRODUCTION: Beta-thalassaemia major is an autosomal recessive disorder that results in severe microcytic, hypochromic, haemolytic anaemia among affected patients. Beta-thalassaemia has emerged as one of the most common public health problems in Malaysia, particularly among Malaysian Chinese and Malays. This study aimed to observe the spectrum of mutations found in Kelantan Malay beta-thalassaemia major patients who attended the Paediatrics Daycare Unit, Hospital Universiti Sains Malaysia, Kelantan, Malaysia, the data of which was being used in establishing the prenatal diagnosis in this Human Genome Centre. METHODS: This was a cross-sectional study conducted with 35 Kelantan Malay beta-thalassaemia major patients. DNA was extracted from the blood collected from the patients and subjected to polymerase chain reaction (PCR) amplification. Six restriction enzymes were used to digest the PCR products for the detection of mutations. RESULTS: Five out of the six beta-globin gene defects were detected, namely, IVS-1 nt5 (G>C), IVS-1 nt1 (G>T), codon 26 (G>A), codon 41-42 (4 bp del) and codon 19 (A>G). The mutation which was not observed in this study was in codon 15 (G>A). The two most common mutations observed were codon 26 (G>A) and IVS-1 nt5 (G>C), which was detected in 26 and 17 patients, respectively. Two patients did not show any of the six mutations. CONCLUSION: Our results added to the existing data on the common beta-globin gene defects in Kelantan Malay beta-thalassaemia patients.

Distribution of the murine plasma cell antigen PC-1 in non-lymphoid tissues.

PC-1 is an alloantigen of murine plasma cells. Its close association with secretory function in lymphoid cells previously raised the question of whether PC-1 was part of the secretory apparatus. In addition to its expression on lymphocytes, PC-1 had been known to be present in liver, brain, and kidney, although the data were derived almost entirely from bulk absorption studies of polyclonal alloantisera, and virtually nothing was known about the nature of the cells expressing PC-1 in these organs. If PC-1 was functionally involved in the secretory process. it might be expected to be present at secretory sites within these and other organs. We now report the results of an immunohistochemical survey of the distribution of PC-1 in a variety of non-lymphoid organs, using a mAb. The PC-1 Ag was found in a small number of highly discrete locations that were mostly, but not exclusively, associated with epithelia. Sites of strong expression included the distal convoluted tubule of the kidney, ducts of the salivary glands, epididymis, proximal part of the vas deferens, and chondrocytes. The PC-1 glycoprotein was also found in the capillaries of the brain, but did not appear to be present in capillaries elsewhere, a pattern that is strikingly similar to that of the receptor for the iron transport protein, transferrin. Negative sites included the thyroid, pancreas, choroid plexus, smooth and striated muscle, stomach, small and large intestine, gall bladder, renal glomeruli, testis, and seminal vesicles. These results are not consistent with a generalized role for PC-1 in secretion, but are compatible with a role in a specialized subset of macromolecular transport events.

The hemoglobin O mutation in Indonesia: distribution and phenotypic expression.

We have investigated hemoglobin O Indonesia (HbOIna) in related ethnic populations of the Indonesian archipelago: 1725 individuals of the five ethnic populations of South Sulawesi (Bugis, Toraja, Makassar, Mandar, and Kajang) and 959 individuals of the neighboring islands, who were divided into five phylogenetic groups: (a) Batak; (b) Malay from Padang, Pakanbaru, and Palembang in the island of Sumatra; (c) Javanese-related populations (Java, Tengger, and Bali) from the islands of Java and Bali; (d) populations of the Lesser Sunda Islands of Lombok, Sumba, and Sumbawa; and (e) the Papuan-languagespeaking population of Alor Island. Nineteen individuals heterozygous for HbO(Ina) were identified from the Bugis, Toraja, Makassar, and Kajang ethnic populations, but none from the other populations. In all cases, the underlying mutation was found to be in codon 116 (GAG to AAG) of the alpha1-globin gene, resulting in the Glull6Lys amino acid change. The level of HbO in the 17 individuals plus 12 additional family members carrying the mutation was found to be 11.6 +/- 1.0%, significantly lower than the expected 17%-22%, indicating the instability of HbO.

Phenotypic variability of Filipino beta(o)-thalassemia/HbE patients in Indonesia.

Three Indonesian patients with identical genotypes, each compound heterozygotes for Filipino beta(o)-thalassemia/HbE, expressed different clinical severities. One patient has mild disease and is transfusion independent, while the other two are severely affected and transfusion dependent. The size of the Filipino beta(o)-globin gene deletion was confirmed to be 45 kb, resolving conflicting values given in the literature. Neither ameliorating genetic factors such as alpha-globin gene deletions or the XmnI restriction site polymorphism at position -158 upstream of the (G)gamma-globin gene, nor differences in beta-globin gene haplotype, explain the phenotypic variation. These observations have implications for the development of antenatal diagnosis in Indonesia, as at present it is not possible to give an accurate prediction of severity of phenotype for this common genotype.