Short stature and an interesting association

Untreated hypothyroidism in children usually results in delayed puberty, but juvenile hypothyroidism causes isosexual precocious puberty in a rare syndrome called Van Wyk Grumbach syndrome, with a complete reversal to the pre pubertal state following thyroid hormone replacement therapy. We report here, a 7-year-old girl who presented with short stature, constipation and isosexual precocious puberty due to the long standing untreated severe hypothyroidism with this syndrome.     

Anti-proliferative and metastatic protease inhibitory activities of protoberberines: An in silico and in vitro approaches

In metastasis, tumour cells interact with numerous factors and one of them is extracellular matrix (ECM). Earlier studies have emphasised the on role of matrix metalloproteases (MMPs) and cathepsin B in ECM degradation and cancer metastasis. In silico docking approaches are a boon in exploring the physicochemical characteristics like angiogenesis, growth and repair of cancerous cells. The binding affinities of berberrubine, jatrorrhizine and thalifendine with caspase 3, MMP-9, cathepsin B and telomeric DNA were performed using hex 6v and iGEMDOCK v2.1 software tools. The present investigation on berberrubine, jatrorrhizine and thalifendine revealed the formation of potentially stable complexes with caspase 3, cathepsin B and telomeric DNA similar to doxorubicin, a well-known anticancer drug. Further, in vitro approaches were made to study anti-proliferative activity against colon, lymphoma and ovarian cancer cell lines and enzyme inhibition activity against MMP-9 and cathepsin B. The results obtained reveal that these protoberberines alkaloids have potential inhibitory effect on cancer cell proliferation as well as metastatic proteases.     

Immunogenicity and protective potential of a bacterially expressed recombinant dihydrolipoamide succinyltransferase (rE2o) of Brucella abortus in BALB/c mice

Brucellosis is one of the world's major zoonoses. No vaccine is available for the prevention of brucellosis in human. Efforts are needed to develop an effective, safe, stable, vaccine with long lasting immunity against human brucellosis. Here, we cloned and expressed recombinant dihydrolipoamide succinyltransferase (rE2o) of Brucella abortus in Escherichia coli and purified up to homogeneity by metal affinity chromatography. The purified rE2o is immunoreactive with brucellosis positive cattle sera. The immunogenicity and the protective potential of recombinant dihydrolipoamide succinyltransferase (rE2o) were evaluated in BALB/c mice with two different adjuvants i.e., Freund's and aluminium hydroxide gel. Mice were tested for humoral immune response by ELISA. Cell mediated immune response was tested by lymphocyte proliferation assay and cytokine profiling. The recombinant E2o (rE2o) generated high IgG antibody and its isotypes IgG1, and induced significant production of INF-γ, IL-10 and IL-4 cytokines. The rE2o protein induced significant lymphoproliferation of splenocytes. Altogether, these results suggest that rE2o induces a mixed but a predominant Th2 type of immune response in BALB/c mice and provides partial protection against challenge with pathogenic Brucella abortus.     

Detection of heteroplasmic mitochondrial DNA in single mitochondria

Background

Mitochondrial DNA (mtDNA) genome mutations can lead to energy and respiratory-related disorders like myoclonic epilepsy with ragged red fiber disease (MERRF), mitochondrial myopathy, encephalopathy, lactic acidosis and stroke (MELAS) syndrome, and Leber''s hereditary optic neuropathy (LHON). It is not well understood what effect the distribution of mutated mtDNA throughout the mitochondrial matrix has on the development of mitochondrial-based disorders. Insight into this complex sub-cellular heterogeneity may further our understanding of the development of mitochondria-related diseases.

Methodology

This work describes a method for isolating individual mitochondria from single cells and performing molecular analysis on that single mitochondrion''s DNA. An optical tweezer extracts a single mitochondrion from a lysed human HL-60 cell. Then a micron-sized femtopipette tip captures the mitochondrion for subsequent analysis. Multiple rounds of conventional DNA amplification and standard sequencing methods enable the detection of a heteroplasmic mixture in the mtDNA from a single mitochondrion.

Significance

Molecular analysis of mtDNA from the individually extracted mitochondrion demonstrates that a heteroplasmy is present in single mitochondria at various ratios consistent with the 50/50 heteroplasmy ratio found in single cells that contain multiple mitochondria.     

Regional Studies of Changes in Brain Fatty Acids Following Experimental Ischaemia and Reperfusion in the Gerbil

Regional studies of brain phospholipid metabolism were carried out during a period of ischaemia induced in the gerbil by bilateral carotid occlusion for 60 min. The associated changes in free fatty acids (FFAs) during this period and following recirculation for up to 180 min were noted. Following ischaemia there was a generalised rise in the levels of all FFAs with no selective release of either the unsaturated (arachidonic and docosahexaenoic) or saturated (palmitic and stearic) fatty acids. There were no observed differences between the brain regions studied, which is in contrast to previously reported observations for prostaglandins. There was also no indication of any specific phospholipid fraction being involved in FFA release. This would indicate that the release of FFAs from phospholipids is a nonspecific event, probably due to the action of hydrolytic lipases. Restoration of the circulation resulted in a short, sharp increase (within 5 min) in all FFAs, but in contrast to the observations during ischaemia alone there was a relatively larger rise in the unsaturated FFAs as compared to the saturated FFAs. Following this increase there was a gradual general decline in all FFA levels until 180 min of reperfusion. Since there was no preferential depletion of unsaturated FFAs during reperfusion, when free radical attack is considered to be at its maximum, it is our opinion that free radical peroxidation is unlikely to explain the pathology described in our model.     

Nonspecific immunoprotection against Rous sarcoma tumor in chicks

An enzyme treated preparation of saprophytic Mycobacterium phlei, referred as NSI, when administered intramuscularly has been found to protect the chicks against Rous Sarcoma Virus induced tumor. A protection level of 35.4%, 24.1% and 21.2% were observed when challenged on 10th, 20th and 30th day post NSI inoculation. The tumor growth inhibitory-activity of NSI was significant (P less than 0.01). Both, systemic and intralesional administration of NSI exhibited significant tumorostatic activity (P less than 0.05). NSI stimulated the cell mediated immune response to specific as well as to nonspecific Rous sarcoma antigen. These studies indicated the immunopreventive activity of NSI against Rous sarcoma tumor which had an immunogenic basis.