High prevalence of West Nile virus in equines from the two provinces of Pakistan.

This study describes the first large-scale serosurvey on West Nile virus (WNV) conducted in the equine population in Pakistan. Sera were collected from 449 equids from two provinces of Pakistan during 2012-2013. Equine serum samples were screened using a commercial ELISA kit detecting antibodies against WNV and related flaviviruses. ELISA-positive samples were further investigated using virus-specific microneutralization tests (MNTs) to identify infections with Japanese encephalitis virus (JEV), WNV and tick-borne encephalitis virus (TBEV). Anti-WNV antibodies were detected in 292 samples by ELISA (seroprevalence 65.0%) and WNV infections were confirmed in 249 animals by MNT. However, there was no animal found infected by JEV or TBEV. The detection of WNV-seropositive equines in Pakistan strongly suggests a widespread circulation of WNV in Pakistan.

Legal framework in aid of biological diversity and statutory deficiencies in Pakistan.

The traditional perception of mutual rights and obligations in the developing world was practically confined to human beings only. Fortunately, this conventional understanding is growing to include all living beings in the scope of certain basic rights. It is also addressing those creatures which are co-existing with human societies naturally or are associated for their subsistence and interdependence. It means that there should be prescribed body of rules and regulations to regulate human conduct in the society. In Pakistan social awakening for animal rights supported by international campaigns have urged state institutions to make adequate body of rules and regulations for protection of animal rights. Purpose of developing required understanding, at the first place is to clarify what is a right and what is an obligation? Secondly what should be the nature of these rights and obligations. Should these be only specific to human beings or there are other beings who also possess certain rights? If other living beings, species have rights, how these rights are to be conferred and regulated? Who is under obligation to provide these rights? Who is liable and responsible to ensure observance of such obligations? It is becoming clearer that like human beings' animals do have certain rights as living beings. All most all religions of the world have prescribed certain rights to them which are encrypted in their holy books. Most of the people are aware of basic concept of observing mild and kind behavior towards pet animals only. Today we scientifically know that humanity and animals have co-existing inevitability. By observing mutual behavior and sensation, the world has provided special rights to the animals by making laws and Acts. Implementation of those rights can only be made through mutual understanding of the citizens under the supervision of law enforcing agencies. It is the responsibility of state and citizens of the states jointly to protect the animals when their rights are being violated.

Chromium-isotope signatures in scleractinian corals from the Rocas Atoll, Tropical South Atlantic.

Chromium-isotope compositions (expressed as δ(53) Cr) of recent and ancient skeletal and non-skeletal carbonates are currently explored as a (paleo-) redox-proxy for shallow seawater. The idea behind this approach is that biogenic and non-biogenic carbonates could potentially be used as archives recording the Cr-isotope composition of seawater in which they formed, and with this contribute to the reconstruction of past paleo-environmental changes in the marine realm, and potentially to climate changes on land. However, investigations addressing the behavior and uptake mechanism of Cr, and the potential isotope fractionations between seawater and biogenic carbonates are scarce. Here, we present a study of Cr-isotope variations in three species of corals and contemporary seawater from the Rocas Atoll, NE, Brazil. Cr-isotope values of the studied coral species (Siderastrea stellata, Porites sp., and Montastrea cavernosa) vary from -0.5 to +0.33‰ and point to significant isotopic disequilibrium with coexisting seawater characterized by a Cr-isotope value of +0.92 ± 0.2‰. This isotopic offset requires reduction of hexavalent Cr(VI) in the sequestration process of all the studied coral species. Cr-isotope values in a profile across an S. stellata colony returned homogeneous, slightly positively fractioned δ(53) Cr values of +0.07 ± 0.08‰ (n = 8, 2σ), which we interpret to reflect a constant reductive uptake during the 20-year growth period recorded in this coral. In contrast, samples across a 12-year growth profile from Porites sp. display rather heterogeneous Cr-isotope values with δ(53) Cr varying from -0.50 to +0.10‰, indicating Cr incorporation under changing redox processes during its growth intervals. We propose a mechanism whereby initial photoreduction of isotopically heavy Cr(VI) to isotopically lighter Cr(III) in the endodermal layer of corals must be followed by efficient and effective re-oxidation of reduced Cr species to favor subsequent chromate (CrO42-) substitution during the calcifying processes ultimately leading to the formation of the coral skeleton.

Sero-epidemiology of equine toxoplasmosis using a latex agglutination test in the three metropolises of Punjab, Pakistan.

Toxoplasmosis is a serious threat for livestock in addition to being of zoonotic significance. In this study, serodiagnosis of equine toxoplasmosis was conducted in a randomly selected population from the 3 metropolises of Punjab, Pakistan. To this end, 272 draught equines were screened using a commercial latex agglutination assay kit. Association of probable risk factors of equine toxoplasmosis was also documented. A total of 91 (33.5%) equines were found sero-positive for Toxoplama (T.) gondii having antibody titers ranging between 1:32 to 1:612. The highest rates of seropositive cases were observed in donkeys (58.7%) followed by mules (28.6%) and horses (23.5%). Age, sex and species of draught equines were found not to be statistically (p>0.05) associated with the distribution of T. gondii antibodies. The results of the study provided a baseline data for the exposure of equine population in this area. In addition, it is recommended that the contiguous population of domestic ruminants and possible reservoirs such as feral cats should be screened in order to explore the potential risk for the human population in Pakistan.

Short communication: guidelines to good manufacturing practices (g.m.p.) in pharmaceutical manufacturing.

The object of the GMP and associated rules is the assurance of the quality of the products for the safety, well being and protection of the patient. In achieving this aim it is impossible to over-emphasise the importance of people, at all levels, in the assurance of the quality of medicinal products. The great majority of reported defective medicinal products has resulted from human error or carelessness and not from failure in technology. All the people involved with the production, quality control or distribution of medicinal products, whether key personnel, production or quality control staff, inspectors or other involved in the many activities which lead to a patient taking a medicine, should bear this constantly in mind when performing their duties.

Differential phototoxicity of fluorescent dye-labeled albumin conjugates.

OBJECTIVE: Fluorescent dyes are commonly used as probes for assessment of macromolecular permeability. Despite numerous examples of light-dye induced toxicity in the microvasculature, little is known regarding the relative phototoxicity of commonly used fluorescent conjugates. We, therefore, compared the phototoxicity of four fluorescent conjugates of bovine serum albumin (BSA) available commercially. METHODS: An in vitro photohemolysis assay was used, in which rat erythrocytes were incubated with fluorescently labeled BSA and exposed to epi-illumination, using an inverted microscope designed for microvascular permeability experiments. Photohemolysis was quantified by monitoring light transmission across the cells. RESULTS: Photohemolysis was dependent on excitation light intensity and fluorochrome concentration. Fluorescein isothiocyanate (FITC)-labeled BSA was the most phototoxic compound tested, inducing 50% of maximum response in 14 min. The relative phototoxicity of the BSA conjugates was: FITC > BODIPY-FL > Texas Red > tetramethylrhodamine isothiocyanate. The phototoxicity of FITC-BSA was related to a high molar dye content. Photohemolysis with each of the conjugates was inhibited by histidine, a singlet oxygen quencher. CONCLUSIONS: Relative phototoxicity of fluorescent albumin conjugates differs considerably. Selection of fluorescent conjugates for use in microvascular experiments based on phototoxicity should consider both the type and molar content of the fluorochrome.

Leakage responses to L-NAME differ with the fluorescent dye used to label albumin.

Nitric oxide synthase (NOS) inhibitors have been reported to increase as well as to decrease microvascular transport of macromolecules in a variety of models. This study was performed to determine whether the influence of NOS inhibition on albumin leakage was dependent on the fluorescent dyes used to label albumin. Albumin leakage was assessed in rat mesenteric venules during control conditions and after exposure to the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME). Albumin was labeled with any one of four dyes: FITC, sulforhodamine 101 [Texas Red (TR)], dichlorotriazinyl aminofluorescein (DTAF), or Oregon Green 514 (OG). Superfusion with L-NAME (10(-4) M) was accompanied by an increase in leakage of FITC-labeled albumin (n = 12) but not of albumin labeled with DTAF (n = 10), TR (n = 10), or OG (n = 4). In vessels perfused with both FITC- and TR-labeled albumin (n = 12), superfusion with L-NAME increased leakage of FITC- but not TR-labeled albumin. In conclusion, albumin leakage responses to L-NAME differ among various fluorescent dyes. Therefore, caution is advised in comparison of albumin leakage results that utilize different fluorescent dyes.

A study of thyrotropin-releasing hormone for the treatment of spinal muscular atrophy: a preliminary report.

OBJECTIVE: To determine whether thyrotropin-releasing hormone (TRH) can increase muscle strength in children with spinal muscular atrophy types 2 and 3. DESIGN: A randomized, double-blinded, controlled, 5-wk drug trial of six subjects and three controls. Subjects and controls ranged from 4 to 8 yr of age and were randomly assigned to treatment and placebo groups in a ratio of 2:1. TRH (protirelin) or placebo was delivered intravenously through percutaneous intravenous catheters at a dose of 0.1 mg/kg (in 50 ml of normal saline) for a total of 29 days. Patients were evaluated using electromyography and handheld dynamometry of the deltoids, biceps, triceps, wrist extensors, hip flexors, quadriceps, hamstrings, and grip strength before and immediately after 5 wk of treatment. A unidirectional t test was used to compare mean values. RESULTS: Dynamometry improved significantly only for the six treated subjects (P < 0.02). Peroneal nerve conduction velocities were significantly faster in the treatment group (paired t test, P = 0.036). The parents of the treated children also provided anecdotal evidence of improvements in function. Improvements lasted 6-12 mo. CONCLUSIONS: TRH may be a useful treatment for spinal muscular atrophy. A larger, crossover design group comparison study is warranted.