Immunomodulatory activity of ethanol extract of Annona reticulata L. leaf in cultured immune cells and in Swiss albino mice.

BACKGROUND: Annona reticulata Linn, has been shown to possess antipyretic, antihelmintic, hypoglycemic, antiulcer and wound healing properties. However, its immunomodulatory role is yet to be explored. OBJECTIVE(S): In the present study, we intended to investigate the effects of A. reticulata leaf ethanol extract on various components of the immune system. MATERIAL AND METHODS: The effects of A. reticulata leaf extract on human peripheral blood mononuclear cells, monocyte (THP1), and human macrophage (U937) cell lines were investigated. An animal study was conducted to observe the effect of the extract on humoral as well as cell mediated immunity. RESULTS: The extract stimulated proliferation of human PBMC, monocytes (THP1), and macrophages (U937) significantly in a dose dependent manner; expression of transforming growth factor-beta (TGF-ß) increased in western blot analysis. Additionally, the extract treated macrophages exhibited features of activation under the microscope with a significant hike in the NO production. Flow cytometry of extract treated human PBMC revealed increased proliferation of lymphocytes (CD4, CD8 & B-cells) along with enhanced intracellular expression of IL-2, IL-6. Animal study data indicate a significant rise in the antibody titer as well as a strong delayed type hypersensitivity response in the extract (150 mg/kg and 300 mg/kg) treated mice; furthermore, the expression of IL-2 and IL-6 in mice PBMC was augmented. CONCLUSION: The collective data evince the immunomodulatory potential of A. reticulata L. leaf.

Evaluation of wound healing activity of ethanol extract of Annona reticulata L. leaf both in vitro and in diabetic mice model.

BACKGROUND AND AIM: The leaves of AnnonareticulataLinn (niú x inguǒ; Bullock's heart), a member of Annonaceae family, have been used extensively in folk medicine; however, its wound healing potential is yet to be explored. Our aim was to investigate the wound healing ability of A. reticulataleaf extract in vitro and in streptozotocin induced diabetic mice model. MATERIAL AND METHODS: We observed the plant extract induced proliferation and migration of primary human dermal fibroblast (HDF), human skin fibroblast cell line (GM00637) and human keratinocyte cell line (HACAT). The expression of transforming growth factor beta (TGF-ß), connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF), alpha smooth muscle actin (α-SMA), matrix metalloproteinases (MMP-2, MMP-9), collagen-1, collagen-3, focal adhesion kinase (FAK) were evaluated by Western blot and gelatin zymography. Excisional diabetic wound model was used for in vivo wound healing assay. Furthermore, we processed wound tissue for histological and immunohistochemical study. RESULT: A. reticulata L. leaf extract stimulates proliferation and migration of HDF, skin fibroblast and keratinocyte significantly in a dose dependent manner; expression of TGF-ß, CTGF, VEGF, α-SMA, MMP-2, MMP-9, collagen-1, collagen-3, FAK increased. Additionally, an enhanced expression of phospho-SMAD2, phospho-SMAD3 in the treated cells indicated the activation of TGF-ß signal transduction pathway, similarly increased expression of phospho-AkT suggested activation of PI3/AkT pathway. Expression of CTGF and α-SMA was also increased significantly in wound tissue. Mass spectrometric analysis revealed that mainly two compounds to be present in the extract: quercetin and ß-sitosterol. CONCLUSION: Collective data suggest that A.reticulata leaf extract may have a stimulatory effect in diabetic wound healing.

Response to Commentary by Adriaan van Es.

I would like to thank Dr Adriaan Van Es for his commentary (1) on my article (2). To start with, let me make one thing clear: I am not sure why he thinks that I am condoning the practice of penal amputation. As I clearly state in my conclusion, the arguments that may (or may not) justify penal amputation are abhorrent in liberal societies. We are on the same side here. But what of those who live in less secular societies where religious faith may be unquestioned? In my opinion, van Es has resorted to a typical example of a tortured form of ethical logic (3), which researchers from countries that have different value systems and different problems have deplored, albeit in a different context.

The ethics of penal amputation.

Malaysia is a South East Asian country with a racially diverse population. Islam is the state religion and about 60% of the population is Muslim, but the rights of other religious groups are protected by law. The Parti Islam se Malaysia, which has ruled the state of Kelantan since 1999, and believes that Malaysia should be ruled by Sharia law, recently proposed the implementation of Hudud laws in Kelantan. However, the federal government has ruled out its implementation. The suggestion stirred up a controversy among the physician community and the Malaysian Medical Association rejected a proposal by the state's political leadership to utilise the services of qualified surgeons to carry out punitive limb amputations. Several Islamic states such as Sudan, Saudi Arabia, and Iran practice Islamic penal justice, including amputations. The question therefore arises: how should a modern medical practitioner approach this ethical question? This study focuses mainly on Malaysia, but draws upon practices in other Islamic countries also.

Ethanolic leaf extract of Coccinia grandis is effective against both drug resistant and drug sensitive clinical isolates of Indian Kala-azar.

The emergence of resistance to the current available drugs used for treatment against Indian Kala-azar (KA) or Visceral Leishmaniasis makes the control strategy inadequate for the disease. This grave epidemiological situation directed researches towards alternative treatments including herbal therapy. In this background, the aim of the present study was to evaluate the antileishmanial activity of the leaves of Coccinia grandis (a tropical vine) against both the Sodium Stibo Gluconate (SSG) sensitive and resistant as well as Miltefosine (MIL) sensitive and resistant field isolates of Leishmania donovani. The cytotoxicity effect of ethanolic extract of leaves of C. grandis (Cg-LE) against the clinical isolates of L. donovani was checked both in promastigotes and intracellular amastigotes stages. In both sensitive and resistant promastigotes, Cg-LE stimulated reactive oxygen species generation and apoptosis. Parasites infected macrophages showing enhanced nitric oxide production after Cg-LE treatment suggested the leishmanicidal activity of the leaf extract. Furthermore, Cg-LE treatment led to mitochondrial membrane damage and DNA fragmentation in promastigotes. The present study is very encouraging for the fact that Cg-LE showed promising antileishmanial activity against both SSG and MIL drug resistant clinical isolates of Indian KA.

Miltefosine Resistant Field Isolate From Indian Kala-Azar Patient Shows Similar Phenotype in Experimental Infection.

Emergence of resistance to drugs used to treat the Indian Kala-azar patients makes control strategy shattered. In this bleak situation, Miltefosine (MIL) was introduced to treat mainly antimonial unresponsive cases. Within years, resistance to MIL has been reported. While checking the MIL sensitivity of the recent KA clinical isolates (n = 26), we came across one isolate which showed four times more EC50 for MIL than that of MIL-Sensitive (MIL-S) isolates and considered as putative MIL-Resistant (MIL-R). The expressions of LdMT and LdRos3 genes of this isolate were found down regulated. Th1/Th2 cytokines, ROS and NO, FACS dot plots and mitochondrial trans membrane potential measurement were performed. In vivo hamster model with this MIL-R isolate showed much lesser reduction in liver weight (17.5%) compared to average reduction in liver weight (40.2%) of the animals infected with MIL-S isolates. The splenic and hepatic stamps smears of MIL-R infected hamsters revealed the retention of parasite load of about 51.45%. The splenocytes of these animals failed to proliferate anti leishmanial T-cells and lack of cell mediated immunity hampered recovery. Thus, these phenotypic expressions of experimental model may be considered similar to that of the MIL unresponsive patients. This is first such kind of report.

Mesenchymal stem cells for cartilage repair in osteoarthritis.

Osteoarthritis (OA) is a degenerative disease of the connective tissue and progresses with age in the older population or develops in young athletes following sports-related injury. The articular cartilage is especially vulnerable to damage and has poor potential for regeneration because of the absence of vasculature within the tissue. Normal load-bearing capacity and biomechanical properties of thinning cartilage are severely compromised during the course of disease progression. Although surgical and pharmaceutical interventions are currently available for treating OA, restoration of normal cartilage function has been difficult to achieve. Since the tissue is composed primarily of chondrocytes distributed in a specialized extracellular matrix bed, bone marrow stromal cells (BMSCs), also known as bone marrow-derived 'mesenchymal stem cells' or 'mesenchymal stromal cells', with inherent chondrogenic differentiation potential appear to be ideally suited for therapeutic use in cartilage regeneration. BMSCs can be easily isolated and massively expanded in culture in an undifferentiated state for therapeutic use. Owing to their potential to modulate local microenvironment via anti-inflammatory and immunosuppressive functions, BMSCs have an additional advantage for allogeneic application. Moreover, by secreting various bioactive soluble factors, BMSCs can protect the cartilage from further tissue destruction and facilitate regeneration of the remaining progenitor cells in situ. This review broadly describes the advances made during the last several years in BMSCs and their therapeutic potential for repairing cartilage damage in OA.

Distal posterior tibial artery pseudoaneurysm after fracture calcaneous.

Here we report a case of pseudoaneurysm of distal posterior tibial artery following closed fracture of calcaneous. Association of pseudoaneurysm of posterior tibial artery with fracture of calcaneous is extremely uncommon. It is always suggested that pseudoaneurysm be treated by reconstruction, but here we present the case treated by ligation, obliteration and excision as it was situated distally and the plantar arch circulation was maintained. The purpose of this case report is to bring attention to this complication of vascular trauma, which may cause delayed symptoms long time after the injury as well as the rare incidence of posterior tibial artery involvement with fracture calcaneous.

P-H and P-P bond activation by Ni(I) and Fe(I) beta-diketiminato-complexes.

Reactions of ((NacNac)Ni)2(mu-eta3-eta3-C6H5Me) (NacNac = HC(CMeNC6H3(i-Pr)2)2) with PH2Ph or PHPh2 proceeds in a facile manner to afford the Ni(I)-phosphine adducts (NacNac)Ni(PH2Ph) and (NacNac)Ni(PHPh2) , respectively. The related reactions of the Fe(I) complex (NacNac)Fe(eta2-CH2CPh2) with PH2Ph resulted in the formation of [(NacNac)Fe(mu2-PPh)]2. Alternatively was also obtained from the reaction of [(NacNac)Fe(mu-Cl)]2 with PHPh2 and Na/K. The Ni(II) di-phosphosphine bridged compound [((NacNac)Ni)2(mu4-(PPh)2)] was prepared via reaction of ((NacNac)Ni)2(mu-eta3-eta3-C6H5Me) with P5Ph5. Crystallographic data are reported for compounds .